|HAI LALA AT MATALIUS009951035B2 DEL TEMA ANA HAITI (12 ) United States Patent (10 ) Patent No. : US 9 , 951, 035 B2 Chang et al. (45 ) Date of Patent: * Apr . 24 , 2018

(54 ) SELECTIVE MATRIX 7 , 928 ,127 B2 4 /2011 Lee et al . 7 ,981 ,426 B2 7 / 2011 Kim METALLOPROTEINASE INHIBITORS 7 , 993 , 665 B2 8 / 2011 Colin et al. 8 , 012 , 947 B2 9 /2011 Tomic et al . @( 71 ) Applicant: University of Notre Dame du Lac, 8 , 093 , 287 B2 1 / 2012 Lee et al . South Bend , IN (US ) 8 , 129 , 341 B2 3 /2012 Gold et al . 8 , 247 , 384 B2 8 / 2012 Green et al . @( 72 ) Inventors : Mayland Chang, Granger , IN ( US ) ; 8 , 889 ,615 B2 11/ 2014 Tomic - Canic et al . 9 ,604 ,957 B2 * 3 / 2017 Chang ...... C07D 331/ 02 Shahriar Mobashery , Granger, IN 2002 / 0037916 A1 3 / 2002 Mobashery et al. (US ) 2007 / 0232541 A1 10 / 2007 Reiter et al. 2009/ 0068251 A1 3 /2009 Tomic et al. @( 73 ) Assignee : University of Notre Dame du Lac , 2011/ 0224275 AL 9 / 2011 Lee et al. South Bend , IN (US ) 2011 / 0293643 Al 12 / 2011 Wilkes 2012 / 0052110 A1 3 /2012 Tomic et al . ( * ) Notice : Subject to any disclaimer , the term of this 2013 / 0052184 Al 2 /2013 Chang et al. patent is extended or adjusted under 35 U . S . C . 154 ( b ) by 0 days. FOREIGN PATENT DOCUMENTS This patent is subject to a terminal dis WO 2006125208 AL 11 /2006 WO 2010093607 AL 8 /2010 claimer . wo 2011026107 AL 3 / 2011 ( 21) Appl. No .: 15 / 469, 010 wo 2016044844 A1 3 / 2016 ( 22 ) Filed : Mar. 24 , 2017 OTHER PUBLICATIONS Armstrong et al. , “ The Role of Matrix Metalloproteinases in Wound (65 ) Prior Publication Data Healing" , Journal of the American Podiatric Medical Association , US 2017 /0190685 A1 Jul. 6 , 2017 vol . 92 , No. 1 , 2002 ; pp . 12 - 18 . Brown et al ., “ Potent and Selective Mechanism - Based Inhibition of Gelatinases” , Journal of the American Chemical Society , vol. 122 , Related U . S . Application Data No . 28 ; Jun . 30 , 2000 ; pp . 6799 -6800 ; American Chemical Society . (63 ) Continuation of application No. 15 / 120 ,508 , filed as Chen et al. , “ Molecular and Mechanistic Validation of Delayed application No . PCT/ US2015 /016950 on Feb . 20 , Healing Rat Wounds as a Model for Human Chronic Wounds ” , Wound Repair and Regeneration , vol. 7 , No. 6 , Nov . 1999 , pp . 2015 , now Pat . No . 9 ,604 , 957 . 486 -494 ; The Wound Healing Society . (60 ) Provisional application No. 62 / 088, 380 , filed on Dec. Cui et al. , “ Inhibition of MMP - 9 by a Selective Gelatinase Inhibitor 5 , 2014 , provisional application No. 61/ 942 , 516 , filed Protects Neurovasculature from Embolic Focal Cerebral Ischemia ” , on Feb . 20 , 2014 . Molecular Neurodegeneration , vol. 7 , No . 21, 2012 , pp . 1 - 15 , BioMed Central Ltd . Fisher et al . , “ Recent Advances in MMP Inhibitor Design ” , Cancer ( 51 ) Int. Cl. and Metastasis Reviews, vol . 25 , 2006 , pp . 115 - 136 , Springer C070 331/ 02 ( 2006 .01 ) Science and Business Media , LLC . CO7D 303 / 34 ( 2006 . 01 ) Forbes et al. , “ Active Site Ring -Opening of a Thiirane Moiety CO7D 413 / 12 ( 2006 .01 ) and Picomolar Inhibition of Gelatinases” , Chemical Biology & A61K 9 /00 ( 2006 . 01 ) Drug Design , vol. 74 , Aug . 2009 , pp . 527 - 534 , John Wiley & Sons A61K 31/ 38 ( 2006 .01 ) A / S . A61K 31/ 4245 ( 2006 .01 ) Gooyit et al. , " A Chemical Biological Strategy to Facilitate Diabetic (52 ) U .S . CI. Wound Healing" , ACS Chemical Biology 2014 , vol . 9 ; Sep . CPC ...... C070 331/ 02 ( 2013 .01 ) ; A61K 9 /0014 20 , 2013 ; pp . 105 - 110 ; ACS Publications, American Chemical (2013 . 01 ) ; A61K 31/ 38 ( 2013 .01 ); A61K Society 31/ 4245 (2013 .01 ) ; C07D 303 / 34 ( 2013 .01 ) ; (Continued ) CO7D 413 /12 (2013 . 01 ) (58 ) Field of Classification Search Primary Examiner — Matthew P Coughlin CPC ...... CO7D 331/ 02 ; CO7D 303 / 34 ( 74 ) Attorney, Agent, or Firm — Haukaas Fortius PLLC ; See application file for complete search history . Michael H . Haukaas ( 56 ) References Cited (57 ) ABSTRACT The invention provides compounds, compositions, and U .S . PATENT DOCUMENTS methods for the treatment of diseases, disorders , or condi 6 , 025 , 150 A 2 / 2000 Livant tions that are modulated by matrix metalloproteinases 6 , 166 ,084 A 12 / 2000 Bloor (MMPs ). The compounds can be selective MMP inhibitors, 6 , 555 , 118 B1 4 / 2003 Niazi for example , selective inhibitors of MMP - 2 , MMP - 9 , and /or 6 , 600 , 057 B2 7 / 2003 Quirk MMP - 14 . The disease, disorder, or condition can include , 6 ,703 , 415 B2 3 / 2004 Mobashery et al. for example , stroke , neurological disorders , ophthalmologi 7 , 144 , 917 B2 12 / 2006 Mobashery et al . 7 , 320 , 783 B2 1 / 2008 Livant cal disorders , or wounds , such as chronic wounds or diabetic 7 ,402 , 571 B2 7 / 2008 Tennenbaum et al. wounds . 7 , 727, 520 B2 6 / 2010 Ferguson 7 , 879 , 798 B1 . 2 /2011 Aufseeser 19 Claims, 9 Drawing Sheets US 9 , 951, 035 B2 Page 2

( 56 ) References Cited Lee et al. , “Metabolism of a Highly Selective Gelatinase Inhibitor Generates Active Metabolite " , Chemical Biology & Drug Design , vol . 70 ; Sep . 2007 , pp . 371 -382 , Blackwell Munksgaard . OTHER PUBLICATIONS Lee et al. , " Structure - Activity Relationship for Thiirane - Based Gooyit et al. , “ O - Phenyl Carbamate and Phenyl Urea Thiiranes as Gelatinase Inhibitors” , ACS Medicinal Chemistry Letters , vol. 3 , Selective Matrix Metalloproteinase - 2 Inhibitors that Cross the May 2 , 2012 ; pp . 490 -495 ; ACS Publications, American Chemical Blood -Brain Barrier” , Journal ofMedicinal Chemistry, vol . 56 , Sep . Society . 12 , 2013 , pp . 8139 - 8150 ; ACS Publications , American Chemical Pradhan et al. , “ Wound -Healing Abnormalities in Diabetes and New Society . Therapeutic Interventions” , Diabetic Foot, US Endocrine Disease , Gooyit et al. , " Selective Gelatinase Inhibitor Neuroprotective 2007 , pp . 68 - 72 , Touch Briefings . Agents Cross the Blood - Brain Barrier " , ACS Chemical Neurosci Song et al . , “ Water - Soluble MMP - 9 Inhibitor Prodrug Generates ence , vol. 3 , Jul. 30 , 2012 ; pp . 730 -736 ; ACS Publications, Ameri Active Metabolites That Cross the Blood - Brain Barrier” , ACS can Chemical Society. Chemical Neuroscience , vol. 4 ; May 20 , 2013 ; pp . 1168 - 1173 ; ACS Gooyit et al. , “ Selective Water- Soluble Gelatinase Inhibitor Publications, American Chemical Society. Prodrugs ” , Journal of Medicinal Chemistry , vol. 54 , Aug . 25 , 2011 ; Toth et al. , “ Tissue Inhibitor of Metalloproteinase ( TIMP) - 2 Acts pp . 6676 -6690 ; ACS Publications, American Chemical Society . Synergistically with Synthetic Matrix Metalloproteinase (MMP ) Gu et al. , “ A Highly Specific Inhibitor of Matrix Metal loproteinase - 9 Rescues Laminin from Proteolysis and Neurons from Inhibitors but not with TIMP -4 to Enhance the (Membrane Type Apoptosis in Transient Focal Cerebral Ischemia ” , The Journal of 1 ) -MMP - DependentActivation of Pro -MMP - 2 ” , The Journal of Neuroscience , vol. 25 , No . 27 , Jul . 6 , 2005 ; pp . 6401- 6408 ; Society Biological Chemistry, vol. 275 , No . 52 , Dec. 29 , 2000 ; pp . 41415 for Neuroscience . 41423 ; The American Society for Biochemistry and Molecular Gutierrez -Fernandez et al. , “ Increased Inflammation Delays Wound Biology , Inc . Healing in Mice Deficient in Collagenase - 2 (MMP - 8 ) " , The FASEB U . S . Patent and Trademark Office as International Searching Journal , vol. 21 , No . 10 , Aug . 2007 , pp . 2580 - 2591 . Authority, “ International Search Report and Written Opinion for Hadass et al. , " Selective Inhibition of Matrix Metalloproteinase - 9 patent application No. PCT /US2015 /016950 , ” dated May 20 , 2015 ; Attenuates Secondary Damage Resulting from Severe Traumatic Brain Injury ” , PLoS One , vol . 8 , No. 10 , e76904 ; Oct. 23 , 2013 , pp . 9 pgs. , published by International Bureau of WIPO (World Intel 1 - 14 ; Open Access. lectual Property Organization ). Harsha et al. , “ ADAM12 : a Potential Target for the Treatment of Zhang et al. , “ Role of Matrix Metalloproteinases and Therapeutic Chronic Wounds” , Journal of Molecular Medicine (Berlin ), vol. 86 , Benefits of Their Inhibition in Spinal Cord Injury ” , No. 8 , Aug . 2008 , pp . 961 - 969, National Institutes of Health Public Neurotherapeutics: The Journal of American Society for Experi Access , 14 pgs . mental NeuroTherapeutics, Apr. 1 , 2011 ; 15 pgs .; Springerlink . com . Hesek et al ., “ Design and Characterization of a Metalloproteinase Zhou et al . , “ QM /MM Studies of the Matrix Metalloproteinase 2 Inhibitor - Tethered Resin for the Detection of Active MMPs in (MMP2 ) Inhibition Mechanism of ( S ) - SB -3CT and Its Oxirane Biological Samples” , Chemistry & Biology, vol. 13 , Apr. 21 , 2006 ; Analogue ” , Journal of Chemical Theory and Computation , vol. 6 , pp . 379 - 386 ; Elsevier Ltd . No . 11 , Jul. 10 , 2010 ; pp . 3580 - 3587 ; American Chemical Society . Hesek et al ., “ Synthesis of an Inhibitor - Tethered Resin for Detection Extended Search Report of the European Patent Office dated Jun . of Active Matrix Metalloproteinases Involved in Disease” , Journal 13, 2017 in EP Application No . 15752642 .7 ; 7pgs. of Organic Chemistry, vol. 71, No . 16 , Jul. 11, 2006 ; pp . 5848 -5854 ; American Chemical Society . * cited by examiner U . S . Patent Apr. 24 , 2018 Sheet 1 of 9 US 9 , 951, 035 B2

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SELECTIVE MATRIX Treatment with the selective gelatinase inhibitor SB - 3CT METALLOPROTEINASE INHIBITORS ( compound 1 ) rescues laminin from proteolysis and neurons from apoptosis following transient focal cerebral ischemia RELATED APPLICATIONS and protected the neurovasculature from embolic focal cere 5 bral ischemia . Disruption of the BBB is observed after This application is a continuation of U . S . patent applica - traumatic brain injury , which has been attributed to MMP - 9 tion Ser. No . 15 / 120 ,508 filed Aug . 19 , 2016 , issued as U . S . and aquaporin -4 (AOP4 ) . Selective inhibition of MMP- 9 Pat. No . 9 , 604 , 957 on Mar. 28 , 2017 , which is a National with compound 1 attenuated secondary damage resulting Stage entry under 35 U . S . C . § 371 of International Patent from severe traumatic brain injury in mice . In mice express Application No. PCT /US2015 /016950 filed Feb . 20 , 2015 , 10 iring mutant superoxide dismutase (SOD 1) that causes amyo which application claims priority under 35 U . S . C . $ 119 ( e ) trophic lateral sclerosis , reduction of MMP - 9 by gene abla to U . S . Provisional Patent Application Nos. 61/ 942 ,516 filed tion , viral gene therapy, or chemical inhibition with the Feb . 20 , 2014 , and 62/ 088 , 380 filed Dec . 5 , 2014 , which tetrapeptidyl hydroxamic acid FN -439 , delayed muscle den applications are incorporated by reference . ervation , indicating that MMP- 9 plays a major role in motor BACKGROUND OF THE INVENTION neuron degeneration . After spinal cord injury , elevated MMP - 9 in the lumbar cord contributes to failure of motor Matrix metalloproteinases (MMPs ) are a group of 26 relearning in mice and deletion of MMP- 9 reduces inflam endoproteases that cleave components of the extracellular mation in the lumbar cord and results in improved recovery . matrix . MMPs exist in their inactive zymogen (proMMP ) 20 MMP - 9 also plays a role in epilepsy. In pentylenetetra forms, requiring activation by disruption of the complex zole - induced epilepsy , sensitivity to epileptogenesis between the cysteine residue in the prodomain and the zinc decreases in mice lacking the MMP - 9 gene and increases in atom in the catalytic domain . MMP activity is regulated rats overexpressing MMP- 9 , and MMP - 9 deficiency dimin predominantly by endogenous inhibitors called tissue inhibi- ishes seizures . MMP- 9 significantly contributes to cell death tors of metalloproteinases ( TIMPs ). MMPs play roles in a 25 after pilocarpine - induced seizures in the developing brain . wide variety of processes , ranging from cell death , differ- Treatment with the broad - spectrum MMP inhibitor GM6001 entiation , proliferation , cell signaling and migration , angio mitigates cell death in pilocarpine - induced seizures in genesis , wound healing, and tissue remodeling . When the immature rats , and in the pathophysiology of brain injury activities of MMPs are imbalanced or uncontrolled , MMPs following seizures. MMP - 1 , MMP- 3 and MMP - 9 have been may play important roles in pathological processes , such as 3030 implicatedm in BBB disruption in West Nile virus encepha tumor metastasis and inflammation . MMPs also play roles in the development and repair of the central nervous system litis . Treatment with the broad - spectrum MMP inhibitor (CNS ) , as well as in the pathology of many neurological GM6001 reversed West Nile virus- induced BBB disruption . diseases . These studies implicate gelatinases in various pathophysi Numerous studies implicating MMPs in cancer pathology 35 Ological processes in the CNS . Thus, selective potent inhibi reresulted in evaluation of broad - spectrum MMP inhibitors in tors of gelatinases that cross the BBB are highly sought. clinical trials in patients with advanced cancer. The com Development of therapeutics that target CNS diseases pounds tested contained hydroxamate groups that chelate to requires that the drugs be delivered to the target site , the zinc , and as such , inhibit MMPs and often other zinc - brain . However, the BBB is a major challenge to the dependent enzymes broadly . Clinical trials with these broad - 40 development of CNS -active small -molecule therapeutics , spectrum MMP inhibitors failed to extend survival. More - constituting a physical barrier that prevents the transport of over , toxic side effects , such as musculoskeletal pain and substances from the blood into the CNS. Small molecules inflammation , were observed . The toxicities were attributed are transported across the BBB by lipid -mediated transport , to the poor selectivity of the inhibitors . In addition , broad if they have a molecular weight of less than 400 Da and / or spectrum MMP inhibitors advanced to clinical trials without 45 high lipid solubility . In practice , a very small number of adequate target validation . It is now recognized that some drugs for CNS diseases fit these criteria . This is due to the MMPs are essential for tumor progression and metastasis , fact that the water solubility of a drug is an important but others play host -protective functions . Thus , the strategy physical property that affects the absorption , distribution , of broad inhibition of MMPs is problematic . Numerous metabolism , and excretion of drugs, as well as whether the studies indicate that neurological conditions would benefit 50 compounds can be screened in a high - throughput manner. from MMP inhibitors , however selective MMP inhibitors While it is assumed that lipophilic small molecules can be are necessary. This is critical, as disparate MMPs mediate transported across the BBB , more than 98 % of small different roles . Some exert the desirable repair functions in molecule drugs do not cross the BBB . Water- soluble drugs disease , yet others promote the deleterious pathological can be lipidized by blocking hydrogen -bond forming func consequences of neurological diseases . 55 tional groups. An example is acetylation of the two hydroxyl A well -studied subgroup of MMPs is the gelatinases: groups in morphine to heroin , which increases BBB pen MMP- 2 ( gelatinase A ) and MMP- 9 ( gelatinase B ) . etration 100 - fold . However, very few CNS drugs have been ProMMP - 2 is present constitutively and is activated by developed by lipidization of water- soluble drugs, as func MMP - 14 , while MMP - 9 is inducible and is activated by tional groups are often metabolized in vivo . Alternatively , a MMP- 3 , plasmin , or under oxidative stress conditions. 60 water- soluble drug may be chemically modified to increase Gelatinases , in particular MMP - 9 , play roles in many patho - its affinity for carrier -mediated BBB transporters . logical CNS conditions, with disruption of the blood - brain Not only must a CNS drug cross the BBB , but it must barrier (BBB ) occurring in many neurological diseases. achieve therapeutic concentrations in the brain and be Following cerebral ischemia , activation of MMP - 2 leads to cleared from the brain so that the drug does not cause CNS disruption of the BBB , followed by a second wave of 65 side effects due to accumulation . As a result , CNS drugs damage to the BBB after reperfusion , which is mediated by have the highest attrition rate in development. Thus, new MMP - 9 . inhibitors of gelatinases that cross the BBB , that achieve US 9 ,951 , 035 B2 therapeutic concentrations in the brain , and that can cleared with respect to the phenoxy moiety of Formula I. In certain from the brain are urgently needed to improve current specific embodiments, R ' is meta or para with respect to the therapies. phenoxy moiety of Formula I. One specific value for J is S . Another specific value for J SUMMARY 5 is O . In one embodiment , G , T , and Q are each independently H . The invention provides selective water - soluble and slow In another embodiment, the compound of Formula I is a binding matrix metalloproteinase inhibitors that cross the compound of Formula II: blood -brain barrier . The matrix metalloproteinase inhibitors can be selective MMP -2 and MMP- 9 inhibitors. These 10 inhibitors can be used for the treatment of neurological ( II) conditions and cancer, as well as for improving the healing of wounds . Accordingly , the invention provides a compound of For- 15ja mula I : RI

(Run wherein R1, R², n , and J are as defined for Formula I . In another specific embodiment, the compound of For mula I or II is : Run O= 25 wherein Rl is CH2 - NHRC wherein Ra is H or ( C . - C . ) alkanoyl; – NH C ( NH ) NH4; or H?N 30

(ND - 336 , 5 ) H?N my * . 35 O HN - S; J is S or O ; og G , T, and Q are each independently H , ( C , - C6) alkyl , or (ND - 378 , 6 ) - CN ; 40 each R2 is independently H , OH , ( C .- C . ) alkyl, (C , -C6 ) alkoxy, (C1 - C . ) alkanoyl, ( C , - C . ) alkanoyloxy, aryl, het eroaryl, carboxy, cyano , nitro , halo , trifluoromethyl , trifluo romethoxy, SR2, SO N (R ) 2, NRR , or COOR ? ; wherein each R * is independently H , (C2 - C . ) alkyl, ( C . - C . ) alkanoylV ., 45 yo ( C6- C10 Jaroyl, aryl, aryl( C . - C . ) alkyl, heteroaryl , heteroaryl ( C - C .) alkyl, or optionally a nitrogen protecting group when R2 is covalently bonded to a nitrogen atom ; and ?? each n is independently 0 , 1 , 2 , 3 , or 4 ; 50 or a salt thereof. HON One value for Rl is — CH - NHRC . O=4 One specific value for Rº is H . ??0 0 Another specific value for R " is C ( O )CH , (acetyl ) . One value for Rl is - NH - C ( = NH ) - NH2. 55 or a pharmaceutically acceptable salt thereof. Another specific value for R is The invention further provides a composition comprising a compound of Formula I or II, in combination with a pharmaceutically acceptable diluent, excipient, or carrier . The pharmaceutical composition can be formulated for HN 60 intravenous , subcutaneous , intracardiac , intramuscular, intraperatoneal, or topical administration . The compounds described herein can be a selective inhibitors of MMP- 2 , MMP - 9 , MMP - 14 , or a combination thereof . For example , the compounds can inhibit MMP - 2 In some embodiments , R is located at the para position 65 and have a K ; of less than about 3 uM . In some embodi with respect to the oxygen of the phenyl group to which R ments, the compound inhibits MMP - 9 and has a K , of less is attached . The variable group R can be ortho ,meta or para than 20 uM . US 9 , 951, 035 B2 The invention also provides a method to inhibit MMP - 2 ing to the patient an effective amount of a compound of or MMP - 9 comprising contacting a composition that Formula I . In some embodiments , the compound of Formula includes MMP - 2 or MMP - 9 with a compound of Formula I, I is a compound of Formula II . In a specific embodiment , the thereby inhibiting the enzymatic activity of the gelatinase . compound is ND - 336 or ND - 378 : In some embodiments , the compound can have an aque - 5 ous solubility of at least about 2 . 5 , at least about 5 , at least about 7 . 5 , at least about 10 , at least about 12 . 5 , at least about ND - 336 15 , at least about 20 , at least about 25 , at least about 30 , or at least about 40 mg/ mL . In some embodiments , the com pound can have an aqueous solubility of at least 2 , 4 , 5 , 10 , 10 20 , 25 , or 30 mM . In yet other embodiments , the compound H2N has an aqueous solubility of at least about 400 times, at least about 4000 times, or at least about 5000 times that of SB -3CT , which has an aqueous solubility of about 2 .45 NDND - 378378 ug/ mL . 15 The invention further provides diagnostic methods and methods of treatment using the compounds described herein , for example, a compound of Formula I. Thus, in one embodiment, the invention provides a method to selectivelywe inhibit a gelatinase comprising contacting the gelatinase 20 with a compound described herein , thereby selectively inhibiting the enzymatic action of the gelatinase . In some embodiments , the inhibition is selective for MMP - 2 in the or a pharmaceutically acceptable salt thereof. presence of MMP - 8 . In some embodiments , the inhibition is In another specific embodiment, the invention provides a selective for MMP - 9 in the presence of MMP - 8 . In some 25 method for a diabetic wound in a patient in need of such embodiments , the inhibition is selective for MMP - 2 in the treatment, the method comprising administering to the presence of both MMP- 9 and MMP - 14 . patient an effective amount of a compound of Formula I . In In some embodiments , the compound is a nanomolar some embodiments, the compound of Formula I is a com slow -binding inhibitor of MMP - 2 , MMP - 9 , and MMP- 14 , pound of Formula II . In a specific embodiment, the com and the compound poorly inhibits MMP- 8 in a non - com - 30 pound is ND - 336 or ND - 378 : petitive manner. In other embodiments , the compound inhib its MMP - 2 , MMP - 8 , MMP- 9 , and MMP - 14 . In various embodiment, the compound inhibits MMP - 2 at a concen ND - 336 tration of less than 0 . 7 uM . In further embodiments , the compound does not inhibit MMP- 1 , MMP- 3 , MMP - 7 , 35 ADAM9, and ADAM10 . The invention also provides a method to treat a neuro H?N logical condition comprising administering to a patient having a neurological condition an effective amount of a V? compound described herein that is effective to treat the 40 ND - 378 neurologicalne condition , thereby alleviating or lessening the symptoms of the condition . The neurological conditions can be one or more of the neurological conditions described herein . The invention also provides a method to treat cancer 45 comprising administering to a patient having cancer an effective amount of a compound described herein that is effective to treat the cancer , thereby killing cancer cells , inhibiting the cancer cells from proliferating , or alleviating or a pharmaceuticallypharmaceutically accepacceptable salt thereof. the symptoms of the cancer. The cancer can be one or more 50 The invention thus provides for the use of the compounds of the cancer conditions described herein . and compositions described herein for use in medical The invention also provides a method to treat a patient therapy . The medical therapy can be treating cancer, for having a wound comprising administering to a patient example , breast cancer, lung cancer, pancreatic cancer, pros having a wound an effective amount of a compound tate cancer, or colon cancer. Themedical therapy can also be described herein that is effective to enhance re - epithelial- 55 therapy for traumatic brain injury or for enhancing the ization of tissues adjacent to the wound . The wound can be healing of wounds . The invention also provides for the use a chronic wound , a diabetic wound , and /or another type of of a compound or composition as described herein for the wound described herein . manufacture of a medicament to treat a condition or disease The invention also provides a method to treat a patient in a mammal, for example, cancer in a human . The medi having traumatic brain injury comprising administering to a 60 ?ament can include a pharmaceutically acceptable diluent, patient having traumatic brain injury an effective amount of excipient, or carrier . a compound described herein that is effective to treat the Accordingly , the invention provides a method of treating traumatic brain injury, thereby lessening the symptoms of a disease or condition that is modulated by a matrix metal the traumatic brain injury. loproteinase (MMP ) comprising administering to a patient in In one specific embodiment, the invention provides a 65 need of such treatment an effective amount of a compound method for treating traumatic brain injury in a patient in of a formula described herein , so that the disease or condi need of such treatment, the method comprising administer - tion is treated . The matrix metalloproteinase (MMP ) can be US 9 ,951 , 035 B2 a gelatinase ( e . g . , MMP - 2 , MMP- 9 , or MMP - 13 ) , a colla neurological disorder or cancer , such as breast cancer, lung genase , a stromelysin , MMP - 23 , MMP- 19 , or matrilysin , cancer, pancreatic cancer, prostate cancer , or colon cancer. and the activity of the matrix metalloproteinase can be Additional diseases , disorders , and conditions that can be significantly inhibited . treated with such therapy are described herein below . The When the compounds, compositions, or methods of the 5 invention also provides for the use of a compound or invention are used to inhibit MMPs, the inhibition may be composition described herein for the manufacture of a selective for one type of MMP over one or more others . In medicament to treat such conditions, for example , condi some embodiments , a compound can selectively inhibit tions in a mammal, such as a human . MMP- 2 , MMP - 9 , and /or MMP- 14 . The manner of inhibition may also involve slow -binding inhibition with respect to kon 10 BRIEF DESCRIPTION OF THE DRAWINGS and koff parameters . Accordingly , modulating a matrix met alloproteinase or inhibiting a matrix metalloproteinase The following drawings form part of the specification and includes selectively inhibiting a matrix metalloproteinase , are included to further demonstrate certain embodiments or such as MMP - 2 , MMP - 9 , and / or MMP - 14 , while other various aspects of the invention . In some instances , embodi gelatinases , such as MMP - 1, MMP- 3 , and / or MMP- 7 are not 15 ments of the invention can be best understood by referring inhibited . to the accompanying drawings in combination with the The disease or condition can include any disease , disor detailed description presented herein . The description and der , or condition recited herein , including , but not limited to , accompanying drawings may highlight a certain specific cancer, stroke , a chronic wound , an ophthalmological dis example , or a certain aspect of the invention . However, one order, traumatic brain injury , spinal cord injury, subarach - 20 skilled in the art will understand that portions of the example noid hemorrhage , tuberculosis , asthma, glaucoma, retinal or aspect may be used in combination with other examples ischemia , ischemic optic neuropathy , macular degeneration , or aspects of the invention . sequalae of hyperhomocystinemia , convulsion , pain , aneu - FIG . 1. Stereo view of the thiirane analogs docked to the rism , depression , anxiety, schizophrenia , muscle spasm , MMP- 2 catalytic site . The inhibitors are represented in migraine headache , emesis , brain edema, tardive dyskinesia , 25 capped sticks, with light gray for carbons, blue for nitrogen , AIDS - induced dementia , ocular damage, retinopathy, a cog - red for oxygen , and yellow for sulfur. Relevant loop amino nitive disorder, or a neuronal injury associated with HIV - acid residues are represented in capped sticks , with purple infection ; or a gelatinase -mediated neurodegenerative dis - for carbon . The zinc ion is shown in gray sphere represen order comprising epilepsy, Alzheimer ' s disease , tation . The Connolly surface was generated for the protein Huntington ' s disease , Parkinson 's disease , multiple sclero - 30 residues excluding that in the loop region , which covers the sis , or amyotrophic lateral sclerosis ; or a combination cavity . Hydrogen bonds between the inhibitor and the pro thereof. tein are shown as black dotted lines . ( A ) The bulky sub In some embodiments , the condition is ischemic stroke or stituent in the terminal ring of inhibitor 4 is tolerated at the hemorrhagic stroke . In another embodiment, the condition is Sl' site ofMMP - 2 . ( B ) Inhibitor 6 , in the absence of steric a neurological disorder or ophthalmological disorder. The 35 hindrance , can form favorable hydrogen bonds with the neurological disorder or ophthalmological disorder can arise backbone carbonyl oxygen atoms of MMP - 2 loop residues . from at least one of trauma, ischemic or hypoxic conditions. The positions of the bulkier residues in other MMPs, Arg424 The neurological disorder can be a neurodegenerative dis - ofMMP - 9 and Gln262 and Met264 ofMMP - 14 , are near the order . In some embodiments , the disease , disorder, or con - 5 o ' clock position of the loop . dition may arise from at least one of painful neuropathy, 40 FIG . 2 . Plasma and brain concentration - time curves in neuropathic pain , diabetic neuropathy , depression , anxiety, mice after single iv dose administration at 5 mg/ kg of ( A ) movement disorders, tardive dyskinesia , cerebral infections compound 4 ; ( B ) compound 5 ; ( C ) compound 6 ; and ( D ) that disrupt the blood - brain barrier , meningitis , meningoen - compound 7 . Concentrations in uM for plasma (triangle data cephalitis , hypoglycemia , cerebral ischemia ( stroke ), car point symbols ) and in pmol/mg tissue for brain ( square data diac arrest, spinal cord trauma, head trauma, perinatal 45 point symbols ) . The x - axis is time in minutes . hypoxia , or hypoglycemic neuronal damage . FIG . 3 . Data from experiments involving a mouse model The administering of a compound described herein can be of diabetic wound healing showing that ND - 336 provides carried out in combination with administering a throm - significantly better efficacy than ND - 322 . Left column = bolytic agent. The thrombolytic agent can be , for example , vehicle ; second from left column = ND - 322 ; second from tissue plasminogen activator ( PA ) . 50 right column = ND - 336 ; right column = ND - 380 . The invention therefore provides novel compounds of FIG . 4 . Picture showing re - epithilialization in the pres Formula I , intermediates for their synthesis , as well as ence of a control ( vehicle ) , ND - 322 , ND - 336 , and ND - 380 . methods of preparing such compounds. The invention also FIG . 5 . A schematic showing that compounds ND - 336 , provides compounds of the formulas described herein that ND - 378 , 4 , and 7 , cross the blood -brain barrier and achieve are useful as intermediates for the synthesis of other useful 55 therapeutic concentrations in the brain . compounds. The invention further provides for the use of FIG . 6 . Gel experiment showing that ND - 336 prevents compounds of Formula I for the manufacture of medica - cleavage of tau in the presence of MMP- 9 . ments useful for the treatment of various conditions modu - FIG . 7 . Inhibition of MMP - 2 has no effect on wound lated by matrix metalloproteinases , such as stroke in a healing in diabetic ( db / db ) mice . Mice were given a single mammal. 60 8 -mm punch biopsy lesion on the skin in the dorsal thorax Additionally , the invention provides compounds and com - under isoflurane anesthesia . One day later , wounds were positions described herein for use in medical therapy . The treated with MMP - 2 inhibitor ( 0 . 25 mg/wound / day ) or invention thus provides for the use of a compound described vehicle ( 20 % DMSO / 80 % propylene glycol) . (a ) Wound herein to prepare a medicament to treat a disease or condi- healing in db / db mice treated with MMP- 2 inhibitor or tion that is modulated by a matrix metalloproteinase (MMP ) . 65 vehicle . Mean + SD ; n = 12 , 6 , and 6 on days 7 , 10 , and 14 , The medicament can include a physiologically acceptable respectively . ( b ) Representative wound images of vehicle diluent or carrier. The medical therapy can be treating a treated and MMP - 2 inhibitor- treated mice . No differences US 9 ,951 , 035 B2 10 were observed between the groups. ( c ) Representative inability of > 98 % of small- molecule drugs to cross the hematoxylin and eosin (H & E ) staining of wounds on day 14 . blood -brain barrier (BBB ) and achieve therapeutic concen Re- epithelialization is indicated by the black line . Scale bars trations in the brain . SB -3CT (compound 1 ) is a selective are 50 m . slow -binding and potent inhibitor of gelatinases that shows FIG . 8 . Ablation of MMP - 9 accelerates wound healing . 5 efficacy in animal models of neurological diseases . How Diabetes was induced by intraperitoneal administration of ever , SB - 3CT is poorly water - soluble . streptozotocin at 150 mg/ kg , as confirmed by measurement We synthesized and evaluated p - aminomethyloxadiazol of fasting blood glucose of > 300 mg/ dL . Wounds were ( 4 ) , p - aminomethyl ( 5 , ND - 336 ) , p - acetamidomethyl ( 6 , inflicted two weeks later . ( a ) Wound healing in MMP - 9 ND - 378 ) , and p - guanidino ( 7 ) as advancements to com knockout and wild - type streptozotocin - induced diabetic 10 Pound 1 . The compounds are 10 - to 14 , 000 - fold more mice. Mean : SD ; n = 14 and 7 on days 7 and 14 , respectively ; water - soluble than 1, retained slow - binding inhibition * p < 0 .05 indicates statistically significant differences in behavior toward MMP- 2 , and crossed the BBB . The p -ac wound healing between MMP - 9 knockout and wild - type etamidomethyl analog ( compound 6 ) is a selective nanomo streptozotocin - induced diabetic mice . ( b ) Representative lar slow - binding inhibitor of MMP- 2 , which does not inhibit wound images ( left, all to the same scale , day 7 ) and H & E 15 the closely related MMP - 9 or MMP - 14 . Because of the slow staining ( right, day 14 ) . Re - epithelialization is indicated by dissociation of compound 6 from the target MMP - 2 (resi the black line . ( c ) In -situ zymography with fluorogenic dence time of 6 bound to MMP- 2 is 18 . 2 + 0 . 4 min ) , it results substrate DQ - gel (DQ - gelatin , green in left panels ) merged in sustained inhibition of MMP - 2 even when concentrations with nuclear DNA staining by DAPI (blue in right panels ) ; of 6 fall below the K , value . This inhibitor is a useful tool in scale bars in panels ( c ) and ( d ) are 50 um . Gelatinase activity 20 therapeutic intervention and in investigations of the role of in wounds was diminished in MMP - 9 knockout mice , as MMP - 2 in neurological diseases . The p - aminomethyl evidenced by significantly decreased fluorescence ( lower derivative ( compound 5 ) is a water- soluble nanomolar slow left ) . binding inhibitor of MMP - 2 , MMP - 9 , and MMP - 14 and has FIG . 9 . Topical treatment with exogenously added residence times for inhibition of these enzymes 6 - to 7 - fold MMP- 8 accelerates wound healing in diabetic (db /db ) mice , 25 longer than those of the tissue inhibitors of metalloprotein and can be combined with the compounds described herein ase 1 or 2 ( TIMP- 1 or TIMP - 2 ) bound to MMP - 9 , protein for using the methods described herein . A single 8 -mm inhibitors that have evolved for the purpose of regulating punch biopsy lesion on the skin in the dorsal thoraxx was MMPs. given to mice under isoflurane anesthesia . Wounds were treated with MMP- 8 ( 1 ug/ wound /day ) or vehicle (saline ). 30 Definitions ( a ) Wound healing in db /db mice treated with MMP - 8 or saline. Mean + SD ; n = 12 , 6 , and 6 on days 7 , 10 , and 14 , The following definitions are included to provide a clear respectively; * p < 0 .05 and # p < 0 .01 indicate statistically sig - and consistent understanding of the specification and claims. nificant differences in wound healing between MMP -8 - As used herein , the recited terms have the following mean treated and vehicle - treated diabetic mice . ( b ) Representative 35 ings. All other terms and phrases used in this specification wound images ( left, all to the same scale ) and H & E staining have their ordinary meanings as one of skill in the art would ( right, day 14 ) . Re - epithelialization is indicated by the black understand . Such ordinary meanings may be obtained by line . ( c ) In - situ zymography with MMP fluorogenic sub - reference to technical dictionaries, such as Hawley ' s Con strate DQ -col I (DQ - collagen , green in left panels ) merged densed Chemical Dictionary 14th Edition , by R . J. Lewis , with nuclear DNA staining by DAPI ( blue in right panels) . 40 John Wiley & Sons, New York , N . Y . , 2001 . Scale bars in panels b and c are 50 um . Topical treatment of References in the specification to " one embodiment” , “ an MMP -8 enhanced the MMP- 8 activity in the wounds, as embodiment” , etc ., indicate that the embodiment described evidenced by the increment of fluorescence (lower left) . may include a particular aspect , feature , structure , moiety , or FIG . 10 . Gel showing the purification of Mus musculus characteristic , but not every embodiment necessarily MMP- 8 . 45 includes that aspect, feature , structure , moiety , or character istic . Moreover, such phrases may , but do not necessarily, DETAILED DESCRIPTION refer to the same embodiment referred to in other portions of the specification . Further , when a particular aspect , feature , The invention may be more fully appreciated by reference structure , moiety, or characteristic is described in connection to the following description , including the following glos- 50 with an embodiment, it is within the knowledge of one sary of terms and the concluding examples . For the sake of skilled in the art to affect or connect such aspect, feature , brevity , the disclosures of the publications , including pat- structure , moiety , or characteristic with other embodiments , ents , cited in this specification are herein incorporated by whether or not explicitly described . reference . Reference is herein made to the subject matter The singular forms “ a , " " an , ” and “ the” include plural recited by certain claims, examples of which are illustrated 55 reference unless the context clearly dictates otherwise . Thus , in the accompanying structures and formulas. While the for example , a reference to " a compound ” includes a plu exemplary subject matter will be described , it will be rality of such compounds, so that a compound X includes a understood that the exemplary descriptions are not intended plurality of compounds X . It is further noted that the claims to limit the claims. On the contrary, the inventive subject may be drafted to exclude any optional element. As such , matter is intended to cover all alternatives , modifications, 60 this statement is intended to serve as antecedent basis for the and equivalents , which may be included within the scope of use of exclusive terminology, such as " solely ," " only, ” and the presently disclosed subject matter as defined by the the like , in connection with any element described herein , claims. and / or the recitation of claim elements or use of “ negative ” Gelatinases (matrix metalloproteinases 2 and 9 ) play limitations. important roles in the pathology of many neurological 65 The term “ and / or” means any one of the items , any diseases. A major challenge to the development of thera - combination of the items, or all of the items with which this peutics for the treatment of neurological diseases is the term is associated . The phrases “ one or more ” and “ at least US 9 ,951 , 035 B2 12 one ” are readily understood by one of skill in the art , embodiments , may be excluded from such categories or particularly when read in context of its usage . For example , embodiments, for example , for use in an explicit negative the phrase can mean one , two , three , four, five, six , ten , 100 , limitation . or any upper limit approximately 10 , 100 , or 1000 times The term “ alkyl” refers to a straight- or branched -chain higher than a recited lower limit . For example , one or more 5 alkyl group having from 1 to about 20 carbon atoms in the substituents on a phenyl ring refers to one to five , or one to chain . For example , the alkyl group can be a ( C , -C20 )alkyl , four, for example if the phenyl ring is disubstituted . a (C2 - C12) alkyl , ( C , - Cg )alkyl , (C , -C .) alkyl, or (C , -C4 ) The term “ about” can refer to a variation of + 5 % , + 10 % , alkyl. Examples of alkyl groups include methyl (Me ) , ethyl + 20 % , or + 25 % of the value specified . For example , " about ( Et) , n - propyl, isopropyl, butyl , isobutyl, sec -butyl , tert 50 ” percent can in some embodiments carry a variation from 10 butyl ( t- Bu ), pentyl, isopentyl, tert -pentyl , hexyl, isohexyl, 45 to 55 percent. For integer ranges , the term “ about ” can and groups that in light of the ordinary skill in the art and the include one or two integers greater than and /or less than a teachings provided herein would be considered equivalent to recited integer at each end of the range . Unless indicated any one of the foregoing examples . Alkyl groups can be otherwise herein , the term “ about” is intended to include optionally substituted or unsubstituted , and optionally par values , e . g ., weight percentages , proximate to the recited 15 tially unsaturated , such as in an alkenyl group . range that are equivalent in terms of the functionality of the The term “ alkenyl ” refers to a straight - or branched - chain individual ingredient, the composition , or the embodiment. alkenyl group having from 2 to 20 carbon atoms in the chain . The term about can also modify the end - points of a recited (The double bond of the alkenyl group is formed by two spa range as discuss above in this paragraph . hybridized carbon atoms. ) Illustrative alkenyl groups As will be understood by the skilled artisan , all numbers , 20 include C , -C12 alkenyl groups , such as prop - 2 - enyl, but- 2 including those expressing quantities of ingredients , prop - enyl, but - 3 -enyl , 2 -methylprop - 2 -enyl , hex - 2 - enyl, and erties such as molecular weight, reaction conditions , and so groups that in light of the ordinary skill in the art and the forth , are approximations and are understood as being teachings provided herein would be considered equivalent to optionally modified in all instances by the term “ about. ” any one of the foregoing examples . Alkenyl groups can be These values can vary depending upon the desired properties 25 optionally substituted or unsubstituted . sought to be obtained by those skilled in the art utilizing the The term " cycloalkyl” refers to a saturated or partially teachings of the descriptions herein . It is also understood saturated , monocyclic , fused polycyclic , or spiro polycyclic that such values inherently contain variability necessarily carbocycle having from 3 to 12 ring atoms per carbocycle , resulting from the standard deviations found in their respec - and can be optionally substituted or unsubstituted . In some tive testing measurements . 30 embodiments , an alkyl group refers to a cycloalkyl group As will be understood by one skilled in the art, for any and that accordingly includes a ring structure . Such alkyl groups all purposes, particularly in terms of providing a written include (cycloalkyl ) - alkyl groups. Illustrative examples of description , all ranges recited herein also encompass any and cycloalkyl groups include the following entities, in the form all possible sub - ranges and combinations of sub - ranges of properly bonded moieties: thereof, aswell as the individual values making up the range , 35 particularly integer values . A recited range (e . g ., weight percentages or carbon groups ) includes each specific value, integer, decimal, or identity within the range . Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least 40 equal halves , thirds , quarters, fifths , or tenths . As a non limiting example , each range discussed herein can be readily DO0. 000 broken down into a lower third ,middle third and upper third , etc . As will also be understood by one skilled in the art, all .000 . language such as " up to " , " at least " , " greater than ” , “ less 45 than ” , “ more than ” , “ or more ” , and the like , include the number recited and such terms refer to ranges that can be subsequently broken down into sub - ranges as discussed ??? above . In the same manner , all ratios recited herein also include all sub -ratios falling within the broader ratio . 50 Accordingly , specific values recited for radicals, substitu ?00 ents , and ranges, are for illustration only ; they do not exclude other defined values or other values within defined ranges for radicals and substituents . One skilled in the art will also readily recognize that 55 7 . D . O . D . where members are grouped together in a common manner, such as in a Markush group , the invention encompasses not only the entire group listed as a whole , but each member of the group individually and all possible subgroups of the , and main group . Additionally , for all purposes, the invention encompasses where the cycloalkyl group is attached at the location of any not only the main group , but also the main group absent one hydrogen atom . or more of the group members . The invention therefore " heterocycle” or “ heterocycloalkyl ” group refers to a envisages the explicit exclusion of any one or more of monocyclic , or fused , bridged , or spiro polycyclic ring members of a recited group . Accordingly, provisos may 65 structure that is saturated or partially saturated and has from apply to any of the disclosed categories or embodiments 3 to 12 ring atoms per ring structure selected from carbon whereby any one or more of the recited elements , species, or atoms and up to three heteroatoms selected from nitrogen , US 9 ,951 , 035 B2 13 14 oxygen , and sulfur. The ring structure may optionally con - heteroaryl can be unsubstituted or optionally substituted . tain up to two oxo groups on carbon or sulfur ring members , Illustrative examples of heteroaryl groups include the fol and can be optionally substituted or unsubstituted . Illustra lowing entities, in the form of properly bonded moieties: tive examples of heterocycle groups include the following entities, in the form of properly bonded moieties: 5

NH 10 N

HN - NH, S N , N , NH : 15 . 1000

NH NH NH 000 20 HN 0

25

HN

?? NH ?? O 30 and Opo EN ' N Droboy- NH , 35 where the heteroaryl group is attached at the location of any hydrogen atom . Those skilled in the art will recognize that the species of cycloalkyl, heterocycle, and heteroaryl groups listed or illustrated above are not exhaustive , and that additional NH , > 40 species within the scope of these defined terms may also be selected . As used herein , the term “ Het” can refer to a 5 or 6 membered heterocyclic ring comprising 1 , 2 , or 3 heteroa COX09 HN toms selected from O , N , S , or P , wherein the ring optionally 45 includes one or two cites of unsaturation and the ring is where the heterocycle group is attached at the location of optionally substituted with 1, 2 , or 3 oxo , halo , nitro , or any hydrogen atom . methyl groups . The Het group can be a heterocycle group or The term “ aryl” refers to an aromatic hydrocarbon group a heteroaryl group . Examples include oxadiazoles , thiadiaz derived from the removal of at least one hydrogen atom from 50 oles, oxazoles , thiazoles, diazines , triazoles , and tetrazoles . a single carbon atom of a parent aromatic ring system . The In one embodiment, Het specifically refers to 1, 3 ,4 - oxadi radical attachment site can be at a saturated or unsaturated azoles, 1 , 2 , 4 -oxadiazoles , the isomeric 1 , 2 , 4 -oxadiazoles , carbon atom of the parent ring system . The aryl group can tetrazoles, 1, 3 ,4 - thiadiazoles, oxazoles, 1 ,2 - diazines, thiaz have from 6 to 30 carbon atoms, for example, about 6 - 14 oles , and 1 , 3 , 4 -triazoles . In another specific embodiment, carbon atoms, about 6 - 13 carbon atoms. or about 6 - 10 55 Het specifically refers to 1 , 2 -diazine , a thiazole , a 1 , 2 , 4 carbon atoms. The aryl group can have a single ring ( e . g . , oxadiazole , a 1 , 3 , 4 - thiadiazole , a 1 , 3 , 4 - triazole , or a tetra phenyl) or multiple condensed ( fused ) rings , wherein at least zole . In yet another embodiment, Het specifically refers to a one ring is aromatic ( e . g . , naphthyl, dihydrophenanthrenyl, 1 , 2 , 4 -oxadiazole , or a 1 , 3 , 4 - thiadiazole . In other embodi fluorenyl, or anthryl) . Typical aryl groups include , but are ments , Het can refer to a 5 -membered heterocyclic ring not limited to , radicals derived from benzene , naphthalene , 60 wherein the ring includes three heteroatoms independently anthracene, biphenyl, and the like . The aryl can be unsub selected from O , S , P , and N . In some embodiments , at least stituted or optionally substituted . two of the heteroatoms are N . In some embodiments , at least The term “ heteroaryl” refers to a monocyclic , fused two of the heteroatoms are O . In yet other embodiments , Het bicyclic , or fused polycyclic aromatic heterocycle ( ring is specifically any 1 , 2 , 3 , 4 , 5 , 6 , 7 , or 8 groups selected from structure having ring atoms selected from carbon atoms and 65 1 ,3 ,4 -oxadiazoles , 1 ,2 ,4 -oxadiazoles , the isomeric 1, 2 ,4 -ox up to four heteroatoms selected from nitrogen , oxygen , and adiazoles, tetrazoles, 1 , 3 , 4 - thiadiazoles , oxazoles , 1 , 2 -diaz sulfur) having from 3 to 12 ring atoms per heterocycle . The ines, thiazoles , and 1 , 3 , 4 - triazoles. US 9 ,951 , 035 B2 15 16 The term “ halogen ” refers to chlorine , fluorine, bromine (C .- C .) alkoxy can be methoxy, ethoxy , propoxy , iso or iodine . The term “ halo ” refers to chloro , fluoro , bromo or propoxy , butoxy , iso -butoxy , sec -butoxy , pentoxy , 3 - pen iodo . toxy, or hexyloxy ; As to any of the groups or “ substituents ” described herein , (C2 - C . )alkenyl can be vinyl, allyl, 1 -propenyl , 2 -prope each can further include one or more (e . g ., 1 , 2 , 3 , 4 , 5 , or 5 nyl, 1 -butenyl , 2 -butenyl , 3 -butenyl , 1 ,- pentenyl , 2 - pente nyl, 3 - pentenyl, 4 -pentenyl , 1 -hexenyl , 2 -hexenyl , 3 - hex 6 ) substituents . It is understood , of course , that such groups enyl, 4 -hexenyl , or 5 -hexenyl ; do not contain any substitution or substitution patterns (C2 - C . ) alkynyl can be ethynyl , 1 - propynyl , 2 - propynyl , which are sterically impractical and /or synthetically non 1 -butynyl , 2 - butynyl, 3 -butynyl , 1 -pentynyl , 2 - pentynyl, feasible . The term “ substituted ” means that a specified group 10 3 -pentynyl , 4 -pentynyl , 1 -hexynyl , 2 -hexynyl , 3- hexynyl , or moiety can bear one or more ( e . g . , 1 , 2 , 3 , 4 , 5 , or 6 ) 4 -hexynyl , or 5 -hexynyl ; substituents . The term " unsubstituted ” means that the speci (C1 - C . )alkanoyl can be acetyl, propanoyl or butanoyl; fied group bears no substituents . The term " optionally sub (C2 - C6) alkanoyloxy can be acetoxy , propanoyloxy , stituted ” means that the specified group is unsubstituted or butanoyloxybu , isobutanoyloxy, pentanoyloxy, or hexanoy substituted by one or more substituents . Where the term 15 lox “ substituted ” is used to describe a structural system , the (C3 - C3) cycloalkyl can be cyclopropyl , cyclobutyl, cyclo substitution is meant to occur at any valency - allowed posi pentyl , cyclohexyl, cycloheptyl , or cyclooctyl; tion on the system . In cases where a specified moiety or aryl can be phenyl, indenyl, 5 , 6 , 7 , 8 - tetrahydronaphthyl, group is not expressly noted as being optionally substituted or naphthyl; and or substituted with any specified substituent, it is understood 20 bicyclic aryl can be indenyl or naphthyl. that such a moiety or group is intended to be unsubstituted Het can be heteroaryl, monocyclic heteroaryl , bicyclic in some embodiments but can be substituted in other heteroaryl, or a non - aromatic heterocycle . Heteroaryl can be embodiments . Suitable substituent groups include , e .g ., furyl, imidazolyl , tetrazolyl, pyridyl (or its N -oxide ) , thie alkyl, alkenyl, alkynyl, alkoxy , halo , haloalkyl, hydroxy , nyl, pyrimidinyl (or its N - oxide ) , indolyl, or quinolyl ( or its hydroxyalkyl, aryl, aroyl, heteroaryl, heterocycle , 25 N - oxide ) ; monocyclic heteroaryl can be furyl, imidazolyl, cycloalkyl, alkanoyl, alkoxycarbonyl, amino , alkylamino , triazolyl, triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiaz dialkylamino , trifluoromethylthio , difluoromethyl, acy oyl, pyrazolyl, pyrrolyl, pyrazinyl, tetrazolyl , pyridyl (or its lamino , nitro , trifluoromethyl, trifluoromethoxy, carboxy, N -oxide ) , thienyl, or pyrimidinyl (or its N -oxide ); and carboxyalkyl, keto , thioxo , alkylthio , alkylsulfinyl, alkylsul- bicyclic heteroaryl can be quinolyl ( or its N -oxide ) ; and fonyl, arylsulfinyl, arylsulfonyl, heteroarylsulfinyl, heteroar - 30 bicyclic alkyl can be decahydroquinoline or decahydronaph ylsulfonyl , heterocyclesulfinyl, heterocyclesulfonyl, phos - thalene (cis or trans) . The Het group can optionally include , phate, sulfate , hydroxyl amine , hydroxyl (alkyl ) amine , and for example , one or two cites of unsaturation , and the ring or cyano . In certain embodiments , any one of the above can optionally be substituted with 1, 2 , 3 , or 4 substituents , groups can be included or excluded from a variable or from for example , oxo , halo , nitro , or methyl groups. a group of substituents . 35 Any formula given herein is intended to represent com Selected substituents within the compounds described pounds having structures depicted by the structural formula herein may be present to a recursive degree . In this context, as well as certain variations or forms. In particular, com " recursive substituent ” means that a substituent may recite pounds of any formula given herein may have asymmetric another instance of itself . Because of the recursive nature of centers and therefore exist in different enantiomeric and/ or such substituents , theoretically , a large number may be 40 diastereomeric forms. All optical isomers and stereoisomers present in any given claim . One of ordinary skill in the art of the compounds of the general formula , and mixtures of medicinal chemistry and organic chemistry understands thereof, are considered within the scope of the formula . that the total number of such substituents is reasonably Thus , any formula given herein is intended to represent a limited by the desired properties of the compound intended . racemate , one or more enantiomeric forms, one or more Such properties include , by of example and not limitation , 45 diastereomeric forms, one or more atropisomeric forms, physical properties such as molecular weight, solubility or and /or mixtures thereof. Furthermore , certain structures may log P , application properties such as activity against the exist as geometric isomers ( i. e . , cis and trans isomers ) , as intended target, and practical properties such as ease of tautomers , or as atropisomers . Additionally , any formula synthesis. In some embodiments , the substitution will result given herein is intended to embrace hydrates , solvates , and in a compound having a molecular weight of less than about 50 polymorphs of such compounds, and mixtures thereof. 1200 Da, less than about 1000 Da, less than about 900 Da, The invention also specifically includes the racemic , less than about 800 Da , less than about 750 Da, less than scalemic , R , and S mixtures and forms at the thiirane moiety about 700 Da, less than about 650 Da, less than about 600 of compounds of Formula A , Formula I , and their associated Da, less than about 500 Da, or less than about 400 Da. formulas. Accordingly , in some embodiments , the stereo Recursive substituents are an intended aspect of the 55 chemistry of the thiirane chiral center is in the R configu invention . One of ordinary skill in the art of medicinal and ration , and in some embodiments , the stereochemistry of the organic chemistry understands the versatility of such sub - thiirane chiral center is in the S configuration . stituents. To the degree that recursive substituents are pres - Compounds of both configurations actively inhibit ent in an embodiment, the total number will be determined MMPs. as set forth above . 60 Any formula given herein is also intended to represent Specific values listed below for substituents (i .e . , groups ) unlabeled forms as well as isotopically labeled forms of the and ranges are for illustration only. They do not exclude compounds. Isotopically labeled compounds have structures other defined values or other values within defined ranges depicted by the formulas given herein except that one or for the substituents. more atoms are replaced by an atom having a selected Specifically , (C , - C .) alkyl can be , for example , methyl, 65 atomic mass or mass number. Examples of isotopes that can ethyl , propyl, isopropyl, butyl , iso -butyl , sec -butyl , pentyl, be incorporated into compounds of the invention include 3 -pentyl , or hexyl; isotopes of hydrogen , carbon , nitrogen , oxygen , phospho US 9 ,951 , 035 B2 rous , fluorine , and chlorine, such as ? H , PH , '1C , 13C , 14C , lowering , stopping or reversing the progression or severity 15N , 180 , 170 , 312 , 32P , 35S, 18F, 36C1 , 1251, respectively . of the condition or symptoms being treated . As such , the Such isotopically labeled compounds are useful in meta - term " treatment" can include medical, therapeutic , and /or bolic studies (preferably with 14C ) , reaction kinetic studies prophylactic administration , as appropriate . ( with , for example ? H or 3H ), detection or imaging tech - 5 The terms “ inhibit ” , “ inhibiting ” , and “ inhibition ” refer to niques ( such as positron emission tomography ( PET) or the slowing , halting, or reversing the growth or progression single -photon emission computed tomography (SPECT )] of a disease , infection , condition , or group of cells . The including drug or substrate tissue distribution assays, or in inhibition can be greater than about 20 % , 40 % , 60 % , 80 % , radioactive treatment of patients . In particular, an 18F or C 10 90 % , 95 % , or 99 % , for example, compared to the growth or labeled compound may be particularly preferred for PET or progression that occurs in the absence of the treatment or SPECT studies . Further, substitution with heavier isotopes contacting . such as deuterium (i . e ., ? H ) may afford certain therapeutic The term “ selective inhibitor ” as used in reference to advantages resulting from greater metabolic stability , for MMPs refers to an inhibitor that inhibits the enzymatic example increased in vivo half - life or reduced dosage 15 activity of one MMP in the presence of one or more other requirements . Isotopically labeled compounds of this inven - MMPs, typically by at least one order of magnitude , for tion and prodrugs thereof can generally be prepared by example , with respect to the Ki. The methods used to obtain carrying out the procedures disclosed in the schemes or in Ki data are known in the art and are described , for example , the examples and preparations described below by substi - , by Brown et al. , J . Amer. Chem . Soc . 2000 , 122 ( 28 ) , 6799 20 6800 , and the references cited therein . Additional useful tuting a readily available isotopically labeled reagent for a assays and techniques are described in U . S . Patent Publica non - isotopically labeled reagent. tion No. 2009/ 0209615 ( Lipton et al. ), which is incorporated When referring to any formula given herein , the selection herein by reference in its entirety . of a particular moiety from a list of possible species for a The term “ mammal” refers to a class of vertebrate animals specified variable is not intended to limit the definition of the 25 of more than 15 ,000 species, including humans , distin moiety for the variable appearing elsewhere. In other words, guished by self- regulating body temperature, hair, and in the where a variable appears more than once , the choice of the females, milk - producing mammae . Mammals include pri species from a specified list is independent of the choice of mates, humans , rodents, canines, felines , bovines, ovines , the species for the same variable elsewhere in the formula , 30 equines, swine , caprines and the like . Specifically , mammal or elsewhere in a different formula . can be a human . Various compounds of Formula I can be readily prepared Selective Water- Soluble and Slow - Binding Matrix Metallo using the techniques described herein , as well as those well proteinase- 2 and - 9 Inhibitors that Cross the Blood - Brain known to those of skill in the art , including the techniques Barrier . described by U . S . Pat. No. 6 , 703 , 415 (Mobashery et al. ) and 35 Evidence is accumulating that damage to neurons and U . S . Pat . No. 7 ,928 , 127 (Lee et al. ) ; PCT Publication No . apoptotic death of neurons play a role in the pathogenesis of WO 2011 /026107 (Mobashery et al. ) ; and U . S . Publication many conditions and disorders , including acute and chronic No. 2013 / 0064878 (Chang et al. ) . neurologic disorders . These disorders range from acute The term “ contacting ” refers to the act of touching , stroke, head trauma, and epilepsy to more chronic conditions making contact, or of bringing to immediate or close prox - 40 such as Huntington ' s disease . Alzheimer ' s disease , HIV imity , including at the cellular or molecular level , for associated dementia , multiple sclerosis , and glaucoma. A example , to bring about a physiological reaction , a chemical contributing factor to several of these diseases is the acti reaction , or a physical change , e . g . , in a solution , in a vation of matrix metalloproteinases (MMPs ) in the extra reaction mixture , in vitro , or in vivo . cellular matrix . An “ effective amount” refers to an amount effective to 45 MMPs constitute a family of extracellular soluble or treat a disease , disorder, and /or condition , or to bring about membrane - bound proteases that are prominently involved in a recited effect. For example , an effective amount can be an remodeling the extracellular matrix . MMP - 9 in particular is amount effective to reduce the progression or severity of the significantly elevated in humans after stroke , which is the condition or symptoms being treated . Determination of a third leading cause ofdeath in the United States . It is also the therapeutically effective amount is well within the capacity 50 primary cause of long - term disability . Acute ischemic of persons skilled in the art . The term " effective amount is stroke, the most common form of stroke , is caused by intended to include an amount of a compound described clotting in the cerebral arteries leading to brain oxygen herein , or an amount of a combination of compounds deprivation and cerebral infarction . Gelatinases ( e . g ., MMP - 2 and MMP - 9 ) are known to be involved in neuronal described herein , e . g ., that is effective to treat or prevent a 55 cell death , blood -brain barrier breakdown and hemorrhage . disease or disorder, or to treat the symptoms of the disease The only FDA -approved drug for the treatment of ischemic or disorder, in a host . Thus , an " effective amount” generally stroke is tissue plasminogen activator (TPA ) , a thrombolytic means an amount that provides the desired effect. agent. The administration of tPA has to be within three hours The terms " treating " , " treat” and “ treatment" can include of the onset of stroke , resulting in its applicability to less ( 1 ) preventing a disease , pathologic or medical condition 60 than 5 % of stroke patients (CNS Neurol. Disord . Drug from occurring ( e . g . , prophylaxis ) ; ( 11 ) inhibiting the disease , Targets 2008 , 7 , 243 - 53 ) . The use of tPA is also limited by pathologic or medical condition or arresting its develop serious side effects , which include neurotoxicity and throm ment; ( iii ) relieving the disease , pathologic or medical bolysis -associated hemorrhagic transformation , and the use condition ; and /or (iv ) diminishing symptoms associated of tPA is contraindicated for patients with evidence of with the disease , pathologic or medical condition . Thus, the 65 hemorrhage or those who are taking anti -coagulant medica terms “ treat” , “ treatment” , and “ treating ” can extend to tion . Blood from stroke patients receiving tPA treatment prophylaxis and can include prevent, prevention , preventing shows elevated levels of MMP - 9 , and tPA was shown to US 9 ,951 ,035 B2 20 activate MMP- 9 . Additionally , recent reports indicate that orders of magnitude more potent against MMP- 2 than tPA upregulates MMP - 9 in the brain and contributes to MMP - 9 and MMP - 14 . The p -aminomethyl analog (com matrix degradation and brain damage . pound 5 ) is a nanomolar slow -binding inhibitor of MMP - 2 , Accordingly , there is a need for new therapies for the MMP - 9, and MMP- 14 , and poorly inhibits MMP -8 in a treatment of stroke, and for treatments of stroke that have 5 nonnon - competitive manner . The p - acetamidomethyl analog fewer and / or less severe side effects than currently used ( compound 6 ) is a selective nanomolar slow - binding inhibi therapies. There is also a need for new gelatinase inhibitors , such as selective gelatinase inhibitors , that do not have the tor of MMP - 2 and does not inhibit the closely related side effects of known therapies such as tPa . MMP- 9 or MMP - 14 . While the p - guanidino derivative The compound SB -3CT (1 ) selectively inhibits gelati 10 ( compound 7 ) is the most potent inhibitor in the series, it nases by a unique mechanism of action involving a reaction lacks selectivity and inhibits MMP - 2 , MMP -8 , MMP - 9 , and catalyzed by the target enzymes , resulting in slow - binding MMP - 14 . and tight -binding inhibition . It is not a metal chelator, as most broad -spectrum metalloprotease inhibitors are . Fur thermore , compound 1 does not broadly inhibit zinc - depen dent proteases, not even other closely - related MMPs. Com pound 1 is rapidly absorbed and readily crosses the BBB , achieving therapeutic concentrations in the brain . However , compound 1 is poorly water -soluble and is metabolized to 20 H2N two major metabolites , with one retaining activity and the other being devoid of it . Our studies on the structure - activity relationships DanND - 336 (5 ) revealed that the terminal ring of the phenoxyphenylmoiety in compound 1 tolerated substitution . One of these inhibi- 25 tors , referred to as ND - 322 (compound 2 ) is water- soluble and inhibits selectively MMP - 2 and MMP - 9 with K , values H2N of 24 and 870 nM , respectively . While compound 2 crosses the BBB , the levels in the brain are below the K , value for MMP- 9 , requiring N - acetylation to the more potent gelati - 30 wou ND- 378 (6 ) nase inhibitor ND - 364 ( compound 3 ) , which achieves thera peutic concentrations in the brain .

SB -3CT ( 1) 35 sayyan 7 NH 40 on HON som ND- 322 (2 ) 45 MMP Inhibition . We evaluated the enzyme kinetics of compounds 4 , 5 , 6 , H?N and 7 with several MMPs and the related ADAM9 ( a 50 disintegrin and metalloproteinase 9 ) and ADAM10 , which our are important enzymes in neurological conditions. Selection of the MMPs for kinetic analysis was based on representa ND - 364 ( 3 ) tive members of the different classes ofMMPs : collagenases 55 (MMP - 1 and MMP - 8 ), gelatinases (MMP - 2 and MMP - 9 ), stromelysins (MMP3 ), matrilysins (MMP - 7 ) , and mem brane - type MMPs (MMP - 14 ) . The results are shown below in Table 1 . All four inhibitors showed potent inhibition in the NH nanomolar range for MMP- 2 and marginal to no inhibition for60 ofMMP -1 , MMP -3 , MMP - 7 , ADAM9, and ADAM10 . The We herein report on the syntheses , MMP kinetics, water p - aminomethyloxadiazol derivative ( compound 4 ) inhibited solubility, pharmacokinetics ( PK ), and brain penetration of MMP - 2 , MMP - 9 , and MMP - 14 in a slow -binding manner four analogs of compound 1 , compounds 4 - 7 . The com with K ; values of 0 .63 + 0 . 06 uM , 34 3 uM , and 9 . 4 + 1 . 1 uM , pounds exhibit 10 - to 14 ,000 - fold increase in water solubil - 65 respectively . A distinctive feature of the thiirane class of ity compared to 1 , retain slow - binding inhibition behavior MMP inhibitors is the slow -binding inhibition of the gelati towards MMP- 2 , and cross the BBB . Compound 4 is 1 - 2 nases . US 9 ,951 ,035 B2 21 TABLE 1 Kinetic Parameters and Aqueous Solubility . K ; (UM ) Enzyme 4 5 Enzyme 4 (p - oxadiazol - 5 (p -CH2NH2 ) 6 (p - 7 (p -guanidino ) 1 ( p- H ) 2 (p -NH2 ) CH2NH2) CHÚNHCOCH3 ) . MMP - 2a 0 .63 + 0 . 06 0 .085 1 0 .001 0 . 23 + 0 .01 0 .021 0 .002 0. 028 + 0 . 0078 0 .024 1 0. 015 MMP- 9a ,b 34 3 0 . 15 + 0 .01 23 % 0 .093 0 . 008 0 .40 0 . 158 0 . 87 + 0 .11 MMP - 14a ,b 9 . 4 + 1 . 1 0 . 12 + 0 . 01 23 % 0 .040 + 0 .002 0 . 11 + 0 .014 , 8 0 .21 + 0 .021 MMP- 15 8 % 4 % 37 % 4 % 73 + 5 , g 10 % , j MMP- 35 17 % 23 % 14 % 19 + 14 4 . 0 + 0 . 44, g 23 .4 1. 61, i MMP- 7 8 % 1 % 5 % 4 % 67 + 61, 8 16 % , j MMP- 86 24 % 7 . 7 + 0 . 1e 0 .69 ± 0 .04 0 .73 + 0 . 056 2 . 1 + 4e 2 .6 + 0 .4e , i ADAMO 5 % 31 % 31 % 14 % 36 % 30 % ADAM10 1 % 14 % 8 % d 25 % 34 % 21 % Aq . solubility 0 . 61 4 . 9 0 .025 32 0 .02338 4 . 9 (mg / mL ) " K ; is calculated from the ratio of kopkom . Slow - binding kinetic parameters ( kon and koffvalues ) are given in Table 2 . " Catalytic domains. Inhibition at 50 UM . dInhibition at 100 uM . Linear noncompetitive mechanism . Linear competitive mechanism . Data from Lee et al. ( Chem . Biol. Drug Des. 2007 , 70, 371 - 382 ); reproduced here for the sake of comparison . " Inhibition at 20 „ M . Data from Gooyit et al. (ACS Chem . Biol. 2014 , 9 , 505 -510 ); reproduced here for the sake of comparison . Inhibition at 250 uM .

Compound 4 was 54 - and 15 - fold more potent against 30 MMP - 9 and Pro253 -Leu271 in MMP - 14 , creating a cavity MMP- 2 than MMP - 9 and MMP - 14, respectively . The that accommodates the terminal ring of the inhibitors . The p - aminomethyl analog ( compound 5 ) was a nanomolar smaller residues Thr426 and Thr428 of the loop line the Si' slow -binding inhibitor of MMP - 2 , MMP -9 , and MMP- 14 site of MMP - 2 , which allows for occupancy by bulkier ( K ; values of 0 .085 + 0 . 001 uM , 0 .015 + 0 .01 uM , and p - substituents on the inhibitors . In contrast , residues Arg424 0 . 12£0 .01 uM , respectively ) ; it poorly inhibited MMP - 8 in ofMMP - 9 and Gln262 and Met264 of MMP - 14 render the a noncompetitive manner (K = 7 .7 + 0 . 1 uM ). The p - acet site less accessible to the larger p - substituents among these amidomethyl derivative ( compound 6 ) was a potent slow - inhibitors . For example , inhibitor 4 with the bulkier p -amin binding inhibitor of MMP- 2 ( K = 0 .23 + 0 .01 uM ) , a linear o methyloxadiazole showed a 54 - fold larger dissociation competitive inhibitor of MMP - 8 ( K ; = 0 .69 + 0 . 04 uM ) , and an constant for MMP - 9 , likely due to the steric clash with poorly inhibited other MMPs and ADAMs, including Arg424 . Arg424 has been reported as a selectivity determi MMP- 9 and MMP - 14 . In contrast, the p -guanidino deriva - nant for MMP - 9 inhibitors in the literature ( Tochowicz et al. , tive (compound 7 ) was the most potent inhibitor in the J . Mol. Biol. 2007 , 371 , 989 - 1006 ). The backbone amide series . However, it lacked selectivity and inhibited MMP - 2 , carbonyl oxygens of the Si ' loop serve as hydrogen bond MMP- 8 , MMP - 9 , and MMP- 14 . 45 donors to substituents on the inhibitors . Docked inhibitor 7 Computational Analysis . with p - guanidino substitution shows multiple potential Our studies on the structural -activity relationship of the hydrogen bonds to the backbone of all of the MMPs , which thiirane class revealed that the sulfonylmethylthiirane moi - can explain why it is the most potent and non - selective ety and phenoxyphenyl group are necessary for inhibition of compound in the series . Inhibitor 5 , with the p - aminomethyl the gelatinases ; these groups are present in compounds 4 - 7 . 50 moiety , also forms similar hydrogen bonds. However , the To rationalize the selectivity of the inhibitors , we carried reason for high selectivity of compound 6 for MMP - 2 was out molecular docking of the compounds to the catalytic not readily evident. It might be reasonable to indicate that sites of MMP - 2 , MMP - 9 , and MMP14 . The caveat to this the steric encumbrance of the Sl' sites of MMP - 9 and analysis is that if a rather large conformational change were MMP - 14 probably renders the p - acetamido moiety less to take place on inhibitor binding, we cannot account for it . » favorable to form hydrogen bonds to the loop backbone . Furthermore, since an X - ray structure for any MMP bound These findings also support the observation that the most to a thiirane inhibitor does not exist presently , we have used potent inhibition by all four compounds was observed for the complex for MMP - 2 bound to the thiirane generated MMP - 2 . based on quantum mechanics /molecular mechanics ( QM ) 60 Compound 5 is 3 - and 6 - fold more potent inhibitor of MM analysis ). However, the results of docking explain a MMP - 9 than compounds 1 and 2, respectively ( Table 1 ). number of features that emerged from the inhibition analy . While compound 4 inhibits MMP - 2 preferentially over ses of these compounds with the panel of MMPs (FIG . 1 ). MMP - 14 ( 15 - fold ), greater than one order of magnitude is The docked poses of the inhibitors show that the p - sub - preferred for therapeutic some in vivo selectivity scenarios ; stituent at the terminal aryl ring fits within the Sl' subsite of 65 however, the inhibition can also be a valuable tool for the MMPs. This subsite is defined by a loop that spans diagnostic analyses . Whereas, compound 6 in vivo will residues Pro417 -Leu433 in MMP - 2 , Pro415 -Leu431 in inhibit MMP - 2 over MMP - 9 ; this compound will also US 9 , 951, 035 B2 23 24 inhibit MMP- 8 to a certain extent. However , for diseases in While compound 5 is 1 . 7 - fold more potent inhibitor of which MMP - 8 does not play a role , compound 6 will MMP- 2 than MMP- 9 , its residence time is 2 - fold longer for binding to MMP - 9 than to MMP - 2 . This indicates that when selectively or exclusively inhibit MMP - 2 . We had previ compound 5 is almost completed eliminated from the brain , ously reported O - phenyl carbamate and phenyl urea thii its slow dissociation from MMP - 9 will result in sustained ranes as selective MMP - 2 inhibitors (Gooyit et al ., J. Med . 5 inhibition of this gelatinasegelatir . Chem . 2013, 56 , 8139 -8150 ), with K , values ranging from Water Solubility . 240 to 760 nM . Compound 6 is a more potent MMP - 2 We determined the water solubility of compounds 4 , 5 , 6 , inhibitor compared to the O - phenyl carbamate and phenyl and 7 by analyzing the filtrate of a saturated aqueous urea thiiranes. Because compound 6 does not inhibit MMP - 9 solution by ultra performance liquid chromatography or MMP - 14 , it is a useful toolbol for ascertaining the rolerole of 10 (UPLC ) with multiple - reaction monitoring (MRM ) . The results are included in Table 1 above . The guanidine analog MMP- 2 in the pathology of diseases . (compound 7 ) is the most water - soluble inhibitor with a One of the most important factors in sustaining efficacy in solubility of 32 mg/ mL . The p - aminomethyl derivative vivo is the drug - target complex residence time, the duration (compound 5 ) is water soluble up to 4 . 9 mg /mL , while the in which the drug is physically bound to the target. The 15 p - aminomethyloxadiazol ( 4 ) water solubility is 0 .61 mg / mL . longer the residence time is , the longer is the duration of The p -acetamidomethyl derivative ( compound 6 ) is the least pharmacological effect. Residence time can be calculated as water soluble among the four, with an aqueous solubility of the reciprocal of the dissociation rate constant (ko ). Resi 0 .025 mg/mL , which corresponds to more than 10 - fold dence times for the inhibitors bound to MMP - 2 , MMP - 9 , improvement in water solubility over compound 1 . Com and MMP- 14 are given in Table 2 . For MMP- 2 , compound 20 pared to compound 2 , addition of a methylene group to give 6 has the shortest residence time of 18 .2 = 0 . 4 min , while compound 5 did not decrease water solubility . compound 4 has the longest at 50 . 5 - 4 . 1 min . The residence The increased water solubility of inhibitors 5 and 7 makes times for all four compounds are longer than those for the them amenable to intravenous (iv ) administration , the pre complexes of MMP - 2 - TIMP -1 or MMP - 2 - TIMP -2 , which ferred route in acute neurological diseases, such as stroke are 7 min and 10 min , respectively (Olson et al. , J. Biol. 25 and traumatic brain injury . Chem . 1997 . 272 . 29975 - 29983 ) . This is an important find - Pharmacokinetics and Brain Distribution . ing , as TIMPs are protein inhibitors of MMPs and have The pharmacokinetics ( PK ) and brain distribution were evolved for inhibition of these enzymes . In essence, these evaluated in mice after single iv dose administration at 5 compounds are more effective in inhibition of the targeted mg/ kg of compounds 4 , 5 , 6 , and 7 . Plasma and brain concentration -time curves and PK parameters are shown in MMPs than are TIMPs. Similarly , the residence times for 30 FIG . 2 and Table 3 , respectively. Plasma concentration of 4 compounds 4 , 5 , and 7 bound to MMP - 9 ranged from 15 . 2 was 62 . 4 : 61 . 6 uM at 2 min , the first time point collected , to 47 . 4 min , which were significantly longer than those for and decreased rapidly to 0 .097 + 0 .09 UM by 30 min , a level MMP- 9 bound to TIMP- 1 or TIMP - 2 of 8 min and 7 min , below the K , value for MMP - 2 . Compound 4 had a moderate respectively , and also longer than that for MMP - 9 bound LOto a35 clearance (CL ) of 0 .0158 L /min /kg ( 19 % of hepatic blood compound 1 of 13 . 4 min . The residence times for com flow ) , a low volume of distribution (Vd ) of 0 .0213 L /kg , a pounds 4 , 5 , and 7 bound to MMP - 14 were similar at very short distribution half - life ( t1 /2Q = 0 .621 min ), and an 14 .6 + 1 . 1 , 12 . 6 + 0 . 3 , and 16 . 4 + 0 .6 , respectively, and were elimination half - life (t1 / 28 ) of 74 . 5 min ( Table 3 ) . Levels of significantly lower than those for TIMP - 2 or TIMP- 4 bound 4 in brainbrain werewere in general lower than those in plasma . Brain to MMP - 14 of 83 min and 39 min , respectively . As the 40 levels of 4 were 9 . 76 + 7 .24 pmol/mg tissue ( equivalent to residence times for the thiirane inhibitors are typically 9 .76 + 7 .24 uM assuming a density of 1 . 0 g /mL ) at 2 min and significantly longer than those for the gelatinase - TIMP decreased below the K , value for MMP - 2 by 30 min . The complexes, the thiirane inhibitors should be equally effec brain to plasma AUCO . ratio was 0 .138 for compound 4 , tive or better in regulating gelatinases. indicating that it crossed the BBB (FIG . 2A , Table 3 ) . TABLE 2 Residence Times for Inhibition of MMPs by the Synthetic Compounds .

MMP- 2 103 koff ( s - ) 0 .330 + 0 . 027 0 .712 + 0 .006 0 .915 + 0 .0190 . 507 + 0 . 049 kon ( s ' M - 1) 521 22 8380 + 110 3910 + 50 2420 + 900 residence time (min ) 50 . 5 4 . 1 23 . 4 0 . 2 18 . 2 + 0 . 4 32 . 9 = 3 . 2 MMP - 9 103 koff (s ) 1 .10 + 0 .03 0 . 352 0. 033 NA 0 .355 + 0 .024 kan ( s - M - 1) 32 .0 + 2 . 3 2360 = 100 NA 3820 + 210 residence time 15 . 2 + 0 . 4 47 . 4 + 4 . 4 NA 47. 0 + 3 . 2 MMP- 14 10% koff (s - ) 1 .14 + 0 . 09 1 . 33 + 0 .03 NA 1 .01 0 .04 kan (s - M - 1) 121 + 11 10800 + 400 NA 25200 + 400 residence time (min ) 14 . 6 1 1. 1 12 . 6 + 0 . 3 NA 16 . 4 + 0 . 6 Not applicable US 9, 951, 035 B2 25 26 TABLE 3 Pharmacokinetic Parameters after Single Intravenous Dose Administration . Compound 4 Compound 5 Compound 6 Compound 7 Parameter Brain Plasma Brain Plasma Brain Plasma Brain Plasma CL ( L /min /Kg ) 0 . 0158 0 . 202 0 .0303 0 . 107 Vd ( L / Kg ) 0 .0213 2 . 76 0 . 329 1 . 08 AUCO- lasia 108108 783783 5757 .. 7 7272 . 9 9 6666 .5 5 435 14 . 3 128128 AUCO. COM 108108 784 58 . 3 3 7373. 6 6767 ..2 437 1717 . 4 128 t1/ 20 (min ) 5 . 35 0 . 621 3 . 32 6 . 47 2 . 89 6 .31 47 5 . 22 t1 /28 (min ) 10 . 9 74 . 5 75 . 3 68 . 0 19 . 1 86 . 6 133 64 . 2 AUC Brain / AUCplasma 0 .138 0 . 793 0 . 154 0 . 136 " AUC in pmol · min /mg for brain and in uMmin for plasma The concentrations of compounds 4 , 5 , 6 and 7 after a 5 mg/ kg single intravenous dose to mice are shown below in Table 4 . TABLE 4 Concentrations of Compounds 4 , 5 , 6 and 7 after 5 mg/ kg Single I. V . Dose to Mice . Time CompoundCompound 4 CompoundCompound 55 (min ) Brain Plasma BrainBrain Plasma 9 . 76 + 7 . 24 62 . 4 + 61 .6 5 . 55 = 2 . 38 4 . 35 + 1 . 10 5 . 98 + 2 . 94 2 . 19 + 0 . 09 2 . 97 2 . 41 3 . 15 1 .49 ÕOUN 3 . 27 + 0 . 14 2 . 14 1 . 51 1 . 20 + 0 . 50 2 . 39 0 . 82 0 . 249 + 0 . 127 0 . 0970 + 0 . 0920 0 . 213 + 0 . 143 0 .377 + 0 .116 60 0 .0253 + 0 .0221 0 .0863 + 0 .0303 0 .0368 + 0 . 0232 0 .0574 + 0 .0316 90 0 .00548 + 0 . 00476 0 .0716 + 0 . 0500 0 .0213 + 0 .0106 0 .0332 = 0 . 0065 120 NDO 0 .0318 + 0 . 0311 0 .0158 + 0 .0125 0 .0218 + 0 . 0054 240 NDCND 0 .0149 = 0 .0129 0 .00531 + 0 .00402 0 .00694 + 0 . 00606 Time Compound 6 Compound 7 (min ) Braina Plasma Brain Plasma 5 . 98 + 1 .48 35 . 0 = 7 . 7 0 .577 + 0 . 560 12 . 6 10 . 9 4 . 90 + 4 . 81 20 . 3 + 2 . 8 0 .476 + 0 .470 4 .37 = 2 .63 0 . 954 + 0 .473 14 . 1 + 5 . 0 0 . 216 + 0 . 071 3 . 92 + 0 . 78 30ON 0 . 231 + 0 . 122 1 .64 + 0 .48 0 . 117 + 0 . 024 0 .482 + 0 . 229 0 . 0544 + 0 .0539 0 .0979 + 0 . 0065 0 .0228 + 0 . 0033 0 . 119 0 .051 90 0 .0263 0 . 0228 0 .0827 + 0 .0262 0 .0409 + 0 .0155 0 .0220 0 . 0080 120 NDC 0 . 0297 + 0 .0168 0 .0222 + 0 .0072 0 . 00630 + 0 . 00490 240 NDO 0 .0193 1 0 .0037 0 .0161 + 0. 0035 0 .00320 0 .00266 " Concentrations in pmol/ mg tissue ; bConcentrations in uM ; Not Detectable .

Plasma and brain concentrations of compound 5 were TABLE 5 above the K , values for MMP - 2 and MMP- 9 for at least 30 Formation of N - Acetyl Metabolite ( Compound 6 ) after min (FIG . 2B and Table 5 ) . As indicated earlier, the slow 55 a 5 mg/kg Single Intravenous Dose of Compound 5 . dissociation of compound 5 from MMP - 9 should result in sustained inhibition even when levels of compound 5 in the Compound 5 _ Compound 6 brain are below the K ; value . Compound 5 had a clearance of 0 . 202 L /min /kg , which is much higher than the hepatic Brain Plasma Brain Plasma AUCO- lasia 57 . 7 72 . 9 0 . 265 0 . 926 blood flow rate of 0 .086 L /min /kg , indicating high clearance AUCO 58. 3 73 .6 NDO 2 . 85 of 5 from systemic circulation with a plasma t1 / 28 of 68. 0 t1 /2a (min ) 3 . 32 6 . 47 4 .05 11. 2 min . Compound 5 had a volume of distribution of 2 . 76 L /kg , t1/ 26 (min ) 75 . 3 68 . 0 NDÓ 990 indicating that 5 was highly distributed to tissues ( Table 3 ) . AUC compound 6/ 5 0 . 0046 0 .039 The brain to plasma AUCO - , ratio of 0 .793 indicated that 65 ( AUC in pmol · min /mg for brain and in uM · min for plasma; compound 5 crossed the BBB readily with a brain t1 / 28 of bNot determined 75 .3 min . US 9 ,951 ,035 B2 27 28 The plasma levels of compound 6 were 35 . 0 + 7 .69 uM at make this compound a useful chemical tool for investigation 2 min and remained above K , for MMP - 2 for 30 min . The of MMP -2 dependent animal models of neurological ail distribution half -life was 6 .31 min and the elimination ments . half - life was 86 .6 min ( FIG . 2C , Table 3 and Table 5 ) . The Conclusions . clearance of compound 6 was 0 . 0303 L /min /kg (35 % of 5 We designed and synthesized four analogs of compound hepatic blood flow rate ), indicating moderate clearance of 6 1 that were 10 - to 14 , 000 - fold more water soluble , retained from systemic circulation (Davies et al ., B . ; Pharm . Res. activity towards MMP - 2 , and crossed the BBB . The p - amin 1993 , 10 , 1093 - 1095 ). The volume of distribution of com omethyloxadiazol analog ( 4 ) is a selective MMP - 2 inhibitor pound 6 was 0 .329 L /kg , indicating that 6 was moderately with a long residence time and moderate water solubility . distributed to tissues ( Table 3 ) . Brain concentrations of 10 The p - acetamidomethyl (compound 6 ) is a selective and compound 6 were 5 . 98 + 1 . 48 pmol/ mg tissue at 2 min and 3 - fold more potent slow -binding inhibitor ofMMP - 2 than 4 ; were above the K ; for MMP - 2 ( 0 . 23 UM , Table 1 ) for 30 min . it does not inhibit MMP - 9 or MMP - 14 . Because of its slow The brain to plasma AUC . .. ratio of 0 . 154 indicated that dissociation from the primary target MMP -2 , the compound compound 6 crossed the BBB and achieved therapeutic 16 can provide sustained inhibition of MMP - 2 even when concentrations in the brain (FIG . 2C , Table 3 and Table 5 ) . concentrations of 6 are below the K ; value . In contrast , The AUC of the p - acetamidomethyl analog ( compound compound 6 inhibits MMP - 8 as a linear competitive inhibi 6 ), an aliphatic amide , relative to that of the p - aminomethyl tor, with a short residence time. Compound 6 crosses the ( compound 5 ) was 0 . 5 % in brain and 3 . 9 % in plasma ( Table BBB and achieves therapeutic concentrations in the brain . 5 ). This indicated minimal in vivo N -acetylation of com - 20 This inhibitor is a useful probe to ascertain the role of pound 5 , an aliphatic amine . In contrast, N -acetylation of the MMP - 2 in neurological diseases . The p -guanidino deriva aromatic amine in compound 2 was significant, as deter tive (compound 7 ) has a water solubility of 32 mg/ mL and mined from the AUC of compound 2 relative to its corre - inhibits MMP - 2 , MMP - 9 , and MMP - 14 in a slow -binding sponding N -acetyl metabolite ( 3 ) of 81 % in brain and 7 . 4 % manner and inhibits MMP - 8 as a linear competitive inhibi in plasma. In addition , the active species after administration 25 tor . The p - aminomethyl derivative ( compound 5 ) is a water of 2 is compound 3 . As the enzymes responsible for N -acety - soluble nanomolar inhibitor ofMMP -2 , MMP - 9 , and MMP lation of compound 2 to compound 3 are polymorphic , direct 14 ; it crosses the BBB and achieves therapeutic administration of compound 3 was required to achieve concentrations in the brain . The residence time of 5 bound therapeutic concentrations in the brain . On the other hand , to MMP - 9 is 6 - to 7 - fold longer than that of TIMP- 1 or p - aminomethyl ( 5 ) is active by itself and does not undergo 30 TIMP- 2 bound to MMP- 9 . Thus , this inhibitor can be significant N -acetylation . equally ormore effective in regulating the activity of MMP The guanidino derivative 7 reached concentrations of 9 . 12 .6 + 10 . 9 uM and 0 .577 + 0 . 560 pmol/mg at 2 min in plasma Therapeutic Methods and brain , respectively . Levels were above the K ; for MMP- 2 A compound of Formula I or a pharmaceutically accept for 90 min in plasma and 120 min in brain , while concen - 35 able salt or solvate thereof ( collectively , " active agents" ) are trations were above the K , for MMP - 9 for 60 min and 30 min useful as MMP inhibitors , or for providing MMP inhibitors in plasma and brain , respectively ( FIG . 2D and Table 5 ). The in vivo , or in vitro , in the methods of the invention . The clearance of compound 7 was high ( 0 . 107 L /min /kg ) and the active agents may be used for the treatment or prevention of high volume of distribution of 1 . 08 L /kg indicated that medical conditions ( such as a wound ) , diseases, or disorders compound 7 was highly distributed to tissues. The plasma 40 mediated through inhibition or modulation of various t128 was 64 . 2 min . The brain to plasma AUC . .. ratio is MMPs, such as those described herein , including the con 0 . 136 , indicating that compound 7 crossed the BBB , with a ditions described in the Background section above . Active brain type of 133 min ( Table 3 ) . The brain of compound agents according to the invention may therefore be used as 7 was significantly longer than those of compounds 4 , 5 , and analgesics, anti -depressants , cognition enhancers, or neuro 6 , suggesting prolonged duration of pharmacological effect. 45 protectants, as well as for treatments for the conditions Comparison of the PK properties of compounds 4 - 7 described below . showed that clearance from plasma was the highest for Described herein are numerous diseases and conditions compound 5 , followed by 7 , 6 , and 4 . Consistent with that might appear to be unrelated but each is related by clearance, plasma systemic exposure (as measured by AUC ) shared mechanistic attributes. Each disease or condition was the highest for compound 4 , then 6 , 7 , and was the 50 described herein is gelatinase - dependent . For example , both lowest for 5 . The volume of distribution followed the same auto -controlled growth and the ability to metastasize are order as clearance : 5 > 7 > 6 > 4 . While the volume of distribu - associated with cancer. The compounds described herein can tion describes the extent that a drug distributes to the body, be anti- proliferative and anti -metastatic toward matrix met it does not imply distribution to the brain . This is because the alloproteinase dependent diseases . endothelial cells of the capillaries of organs allow passage of 55 Compounds and pharmaceutical compositions suitable for small molecules from blood into organs, however those of use in the invention include those wherein the active agent the brain capillaries are surrounded by tight junctions that is administered in an effective amount to achieve its intended prevent drugs in circulation from entering the brain . As a purpose . The phrase " therapeutically effective amount" result, the volume of distribution does not correlate with the refers to an amount effective to treat the disease, disorder, AUChroin blood , which was highest for compound 5 > 6 > 4 = 7 . 60 and /or condition , for example , an amount effective to reduce The highbrain /bloo potency for MMP - 9 , the long residence time for the progression or severity of the condition or symptoms inhibition ofMMP - 9 , along with distribution of compound being treated . Determination of a therapeutically effective 5 to the brain and clearance from the brain , make compound amount is well within the capacity of persons skilled in the 5 worthy of administration in animal models of MMP - 9 art . The term " effective amount" can include an amount of dependent neurological diseases . Likewise , the potency and 65 a compound described herein , or an amount of a combina selectivity of compound 6 for MMP- 2 and its ability to cross tion of compounds described herein , e . g ., to treat or prevent the BBB and achieve therapeutic concentrations in the brain a disease or disorder , or to treat the symptoms of the disease US 9 , 951, 035 B2 29 30 or disorder , in a host . Thus , an “ effective amount" generally a pharmaceutically acceptable diluent or carrier. The phar means an amount that provides the desired effect. maceutical composition can include a thrombolytic agent or Exemplary medical conditions , diseases , and disorders an analgesic , such as an opioid or a non - steroidal anti include anxiety, depression , pain , sleep disorders , inflam inflammatory drug . Examples of such analgesics include mation , multiple sclerosis and other movement disorders , 5 aspirin , acetaminophen , opioids , ibuprofen , naproxen , HIV wasting syndrome, closed head injury , stroke , learning COX - 2 inhibitors , gabapentin , pregabalin , tramadol, or and memory disorders , Alzheimer ' s disease , epilepsy, combinations thereof. Tourette 's syndrome, epilepsy, Niemann -Pick disease , Par The term “ thrombolytic agent” refers to a drug that is able kinson ' s disease , Huntington ' s chorea , optic neuritis , auto - to dissolve a clot or “ thrombus” and reopen an artery or vein . immune uveitis , symptoms of drug withdrawal, nausea , 10 Thrombolytic agents may be used to treat a heart attack , emesis , sexual dysfunction , post - traumatic stress disorder, stroke , deep vein thrombosis ( e . g ., a clot in a deep leg vein ) , or cerebral vasospasm , or combinations thereof, as well as pulmonary embolism , and occlusion of a peripheral artery or the conditions described below . indwelling catheter. Thrombolytic agents are serine pro The active agents may be used to treat subjects (patients ) teases and they convert plasminogen to plasmin , which diagnosed with or suffering from a disease , disorder , or 15 breaks down fibrinogen and fibrin and dissolves blood clots . condition that is mediated through MMP activity , e . g . , one of Currently available thrombolyic agents include reteplase the 26 known gelatinases . The term “ treat” or “ treating ” as ( r - PA or Retavase ) , alteplase ( t - PA or Activase ) , urokinase used herein is intended to refer to administration of an agent ( Abbokinase ) , prourokinase, anisoylated purified streptoki or composition of the invention to a subject for the purpose nase activator complex (APSAC ) , and streptokinase . of effecting a therapeutic or prophylactic benefit through 20 Thrombolytic agents are also called clotbusters, clot -dis modulation of MMP activity . Treating includes reversing , solving medications, and fibrinolyic agents . ameliorating , alleviating , inhibiting the progress of, lessen Accordingly , the invention also provides a method of ing the severity of, or preventing a disease , disorder , or treating a subject suffering from or diagnosed with a disease , condition , or one or more symptoms of such disease , dis - disorder, or medical condition mediated by MMP activity , order or condition mediated through modulation of MMP 25 comprising administering to the subject in need of such activity . treatment an effective amount of at least one compound of The term “ subject” refers to a mammalian patient in need a formula described herein , a pharmaceutically acceptable of such treatment, such as a human . “ Modulators ” include salt thereof, a pharmaceutically acceptable prodrug thereof, both inhibitors and activators, where “ inhibitors ” refer to or a pharmaceutically active metabolite thereof. The disease , compounds that decrease , prevent, inactivate , desensitize or 30 disorder, or medical condition can include anxiety , depres down -regulate MMP expression or activity , and “ activators” sion , pain , sleep disorders, eating disorders , inflammation , are compounds that increase , activate , facilitate , sensitize , or movement disorders , HIV wasting syndrome, closed head up - regulate MMP expression or activity . injury, stroke , Alzheimer ' s disease, epilepsy, Tourette ' s syn Accordingly, the invention relates to methods of using the drome, Niemann -Pick disease , Parkinson ' s disease , Hun active agents described herein to treat subjects diagnosed 35 tington ' s chorea , optic neuritis , autoimmune uveitis , drug with or suffering from a disease , disorder , or condition withdrawal, nausea , emesis , post- traumatic stress disorder, mediated through MMP activity , such as anxiety , pain , sleep cerebral vasospasm , glaucoma, irritable bowel syndrome, disorders, inflammation , or movement disorders ( e . g . , mul- inflammatory bowel disease , immunosuppression , gastroe tiple sclerosis ). sophageal reflux disease, paralytic ileus , secretory diarrhea , Symptoms or disease states are intended to be included 40 gastric ulcer , rheumatoid arthritis , hypertension , cancer, within the scope of “ medical conditions , disorders , or dis - hepatitis, allergic airway disease , autoimmune diabetes , eases . " For example , pain may be associated with various intractable pruritus , neuroinflammation , or a combination diseases , disorders , or conditions , and may include various thereof . etiologies . Illustrative types of pain treatable with a MMP The invention further includes a pharmaceutical compo modulating agent according to the invention include cancer 45 sition for treating a disease , disorder , or medical condition pain , postoperative pain , GI tract pain , spinal cord injury mediated by MMP activity , comprising: ( a ) an effective pain , visceral hyperalgesia , thalamic pain , headache (includ amount of at least one compound of a formula described ing stress headache and migraine ) , low back pain , neck pain , herein , or a pharmaceutically acceptable salt , a pharmaceu musculoskeletal pain , peripheral neuropathic pain , central tically acceptable prodrug, or a pharmaceutically active neuropathic pain , neurogenerative disorder related pain , and 50 metabolite thereof, or any combination thereof, and a phar menstrual pain . HIV wasting syndrome includes associated maceutically acceptable excipient. symptoms such as appetite loss and nausea . Parkinson ' s Diseases , Disorders , and Conditions disease includes, for example , levodopa - induced dyskinesia . The compounds and compositions described herein can be Treatment of multiple sclerosis may include treatment of used to treat or reduce the symptoms of the following symptoms such as spasticity , neurogenic pain , central pain , 55 diseases , disorders , and conditions. or bladder dysfunction . Symptoms of drug withdrawal may The term “ neurological disorder ” refers to any disorder of be caused by, for example , addiction to opiates or nicotine . the nervous system and /or visual system . " Neurological Nausea or emesis may be due to chemotherapy , postopera - disorders” include disorders that involve the central nervous tive , or opioid related causes. Treatment of cancer may system (brain , brainstem and cerebellum ) , the peripheral include treatment of glioma. Sleep disorders include , for 60 nervous system ( including cranial nerves ) , and the auto example , sleep apnea , insomnia , and disorders calling for nomic nervous system (parts of which are located in both treatment with an agent having a sedative or narcotic -type central and peripheral nervous system ) . Neurodegenerative effect . Eating disorders include, for example , anorexia or disorder also refers to a type of neurological disease marked appetite loss associated with a disease such as cancer or HIV by the loss of nerve cells , including , but not limited to , infection / AIDS . 65 Alzheimer ' s disease , Parkinson ' s disease, amyotrophic lat The invention also provides a composition comprising a eral sclerosis , tauopathies ( including fronto - temporal compound of any one of the formulas described herein and dementia ), and Huntington 's disease . US 9 , 951, 035 B2 31 32 Major groups of neurological disorders include, but are ease; Kugelberg - Welander disease ; Kuru ; Lafora disease ; not limited to , headache , stupor and coma, dementia , sei- Lambert - Eaton myasthenic syndrome; Landau -Kleffner syn zurezu , sleep disorders, trauma, infections, neoplasms, neu drome; Lateral medullary (Wallenberg ) syndrome; Learning roophthalmology, movement disorders, demyelinating dis disabilities; Leigh ' s disease; Lennox -Gastaut syndrome; eases, spinal cord disorders , and disorders of peripheral 5 Lesch -Nyhan syndrome: Leukodystrophy : Lewy body nerves, muscle and neuromuscular junctions. Addiction and dementia : Lissencephaly : Locked - in syndrome: Lou Geh mental illness , include , but are not limited to , bipolar dis order and schizophrenia , are also included in the definition rig ' s disease (aka Motor Neuron Disease or Amyotrophic of neurological disorder. The following is a list of several Lateral Sclerosis ) ; Lumbar disc disease; Lyme disease neurological disorders , symptoms, signs and syndromes: 10 Neurological Sequelae ; Machado - Joseph disease; Macren Acquired Epileptiform Aphasia ; Acute Disseminated cephaly ; Megalencephaly ; Melkersson -Rosenthal syn Encephalomyelitis ; Adrenoleukodystrophy; Agenesis of the drome; Menieres disease ; Meningitis ; Menkes disease ; corpus callosum ; Agnosia ; Aicardi syndrome; Alexander Metachromatic leukodystrophy ; Microcephaly ; Migraine ; disease; Alpers' disease ; Alternating hemiplegia ; Alzheim Miller Fisher syndrome; Mini- Strokes; Mitochondrial Myo er ' s disease ; Amyotrophic lateral sclerosis ; Anencephaly ; 155 pathiespathi ; Mobius syndrome; Monomelic amyotrophy ; Motor Angelman syndrome; Angiomatosis ; Anoxia ; Aphasia ; Neurone Disease ; Moyamoya disease ; Mucopolysacchari Apraxia ; Arachnoid Cysts ; Arachnoiditis ; Amold - Chiari doses; Multi - Infarct Dementia ; Multifocal motor neuropa malformation ; Arteriovenous malformation ; Asperger syn thy ; Multiple sclerosis and other demyelinating disorders ; drome; Ataxia Telangiectasia ; Attention Deficit Hyperactiv Multiple system atrophy with postural hypotension ; Mus ity Disorder ; Autism ; Autonomic Dysfunction ; Back Pain ; 20 cular dystrophy ; Myasthenia gravis ; Myelinoclastic diffuse Batten disease ; Behcet' s disease ; Bell' s palsy ; Benign sclerosis ; Myoclonic encephalopathy of infants ; Myoclonus; Essential Blepharospasm ; Benign Focal ; Amyotrophy ; Myopathy ; Myotonia congenital; Narcolepsy ; Neurofibro Benign Intracranial Hypertension ; Binswanger 's disease ; matosis ; Neuroleptic malignant syndrome; Neurological Blepharospasm ; Bloch Sulzberger syndrome; Brachial manifestations of AIDS ; Neurological sequelae of lupus; plexus injury ; Brain abscess ; Brain injury ; Brain tumors 25 Neuromyotonia ; Neuronal ceroid lipofuscinosis ; Neuronal ( including Glioblastomamultiforme ) ; Spinal tumor; Brown - migration disorders ; Niemann - Pick disease ; O 'Sullivan Sequard syndrome; Canavan disease ; Carpal tunnel syn McLeod syndrome; Occipital Neuralgia ; Occult Spinal Dys drome (CTS ); Causalgia ; Central pain syndrome; Central raphism Sequence ; Ohtahara syndrome ; Olivopontocerebel pontine myelinolysis ; Cephalic disorder; Cerebral aneu lar Atrophy ; Opsoclonus Myoclonus; Optic neuritis ; rysm ; Cerebral arteriosclerosis ; Cerebral atrophy ; Cerebral 30 Ort gigantism ; Cerebral palsy ; Charcot- Marie - Tooth disease ; Orthostatic Hypotension ; Overuse syndrome; Paresthesia ; Chemotherapy - induced neuropathy and neuropathic pain ; Parkinson ' s disease ; Paramyotonia Congenita ; Paraneoplas Chiari malformation ; Chorea ; Chronic inflammatory demy tic diseases ; Paroxysmal attacks; Parry Romberg syndrome; elinating polyneuropathy ( CIDP ) ; Chronic pain ; Chronic Pelizaeus- Merzbacher disease ; Periodic Paralyses ; Periph regional pain syndrome; Coffin Lowry syndrome; Coma, 35 eral Neuropathy ; Painful Neuropathy and Neuropathic Pain ; including Persistent Vegetative State ; Congenital facial Persistent Vegetative State ; Pervasive developmental disor diplegia ; Corticobasal degeneration ; Cranial arteritis ; Cran ders ; Photic sneeze reflex ; Phytanic Acid Storage disease ; iosynostosis ; Creutzfeldt- Jakob disease ; Cumulative trauma Pick ' s disease ; Pinched Nerve ; Pituitary Tumors ; Polymyo disorders; Cushing ' s syndrome: Cytomegalic inclusion body sitis ; Porencephaly ; Post - Polio syndrome; Postherpetic Neu disease (CIBD ) ; Cytomegalovirus Infection ; Dancing eyes - 40 ralgia (PHN ) ; Postinfectious Encephalomyelitis ; Postural dancing feet syndrome; Dandy - Walker syndrome; Dawson Hypotension ; Prader - Willi syndrome; Primary Lateral Scle disease; De Morsier' s syndrome; Dejerine - Klumpke palsy ; rosis ; Prion diseases ; Progressive ; Hemifacial Atrophy ; Pro Dementia ; Dermatomyositis ; Diabetic neuropathy ; Diffuse gressive multifocal leukoencephalopathy ; Progressive Scle sclerosis ; Dysautonomia ; Dysgraphia ; Dyslexia ; Dystonias ; rosing Poliodystrophy ; Progressive Supranuclear Palsy ; Early infantile epileptic encephalopathy ; Empty sella syn - 45 Pseudotumor cerebri; Ramsay -Hunt syndrome ( Type I and drome; Encephalitis ; Encephaloceles ; Encephalotrigeminal Type II ) ; Rasmussen ' s Encephalitis ; Reflex Sympathetic angiomatosis ; Epilepsy ; Erb ' s palsy ; Essential tremor; Fab Dystrophy syndrome; Refsum disease ; Repetitive Motion ry ' s disease ; Fahr' s syndrome; Fainting ; Familial spastic Disorders ; Repetitive Stress Injuries ; Restless Legs syn paralysis ; Febrile seizures ; Fisher syndrome; Friedreich ' s drome; Retrovirus - Associated Myelopathy ; Rett syndrome; ataxia ; Fronto - Temporal Dementia and other " Tauopathies” ; 50 Reye ' s syndrome ; Saint Vitus Dance ; Sandhoff disease ; Gaucher ' s disease ; Gerstmann 's syndrome; Giant cell arteri- Schilder ' s disease ; Schizencephaly ; Septo - Optic Dysplasia ; tis ; Giant cell inclusion disease ; Globoid cell Leukodystro - Shaken Baby syndrome; Shingles ; Shy -Drager syndrome; phy; Guillain - Barre syndrome; HTLV - 1 associated Sjogren 's syndrome; Sleep Apnea ; Soto 's syndrome; Spas myelopathy ; Hallervorden - Spatz disease ; Head injury ; ticity ; Spina bifida ; Spinal cord injury ; Spinal cord tumors ; Headache ; Hemifacial Spasm ; Hereditary Spastic Paraple - 55 Spinal Muscular Atrophy ; Stiff -Person syndrome; Stroke ; gia ; Heredopathia atactica polyneuritiformis ; Herpes zoster Sturge -Weber syndrome; Subacute Sclerosing Panencepha oticus; Herpes zoster; Hirayama syndrome; HIV -Associated litis ; Subarachnoid Hemorrhage ; Subcortical Arterioscle Dementia and Neuropathy (see also Neurological manifes- rotic Encephalopathy ; Sydenham Chorea ; Syncope ; Syrin tations of AIDS) ; Holoprosencephaly ; Huntington ' s disease gomyelia ; Tardive dyskinesia; Tay - Sachs disease ; Temporal and other polyglutamine repeat diseases; Hydranencephaly ; 60 arteritis ; Tethered Spinal Cord syndrome; Thomsen disease ; Hydrocephalus ; Hypercortisolism ; Hypoxia ; Immune -Me - Thoracic Outlet syndrome; Tic Douloureux , Todd ' s Paraly diated encephalomyelitis ; Inclusion body myositis ; Inconti- sis ; Tourette syndrome; Transient ischemic attack ; Trans nentia pigmenti ; Infantile ; phytanic acid storage disease ; missible Spongiform Encephalopathies; Transverse myeli Infantile Refsum disease ; Infantile spasms; Inflammatory t is ; Traumatic Brain injury ; Tremor , Trigeminal Neuralgia ; myopathy ; Intracranial cyst ; Intracranial hypertension ; Jou - 65 Tropical Spastic Paraparesis ; Tuberous Sclerosis ; Vascular bert syndrome; Kearns - Sayre syndrome ; Kennedy disease ; Dementia (Multi -Infarct Dementia ) ; Vasculitis including Kinsbourne syndrome; Klippel Feil syndrome; Krabbe dis - Temporal Arteritis ; Von Hippel -Lindau Disease (VHL ); US 9 ,951 , 035 B2 33 34 Wallenberg ' s syndrome; Werdnig -Hoffman disease; West nocerebellar ataxia type 3 ) , Multiple System Atrophy, syndrome; Whiplash ; Williams syndrome; Wilson ' s disease ; Pelizaeus -Merzbacher disease , Pick ' s disease , primary lat and Zellweger syndrome. eral sclerosis , Refsum 's disease , Sandhoff disease , Schil The medical therapy can also be for the treatment of der ' s disease, spinocerebellar ataxia (multiple types with cancer , angiogenesis , cardiovascular disease , neurological 5 varying characteristics ), spinal muscular atrophy , Steele disease , inflammation , eye disease , autoimmune disease , or Richardson - Olszewski disease, or tabes dorsalis . other conditions that are affected by the regulation ofMMPs . The compounds described herein can be used to treat The cancer can be pancreatic cancer, gastric cancer , lung conditions of the eye , including corneal wounds , glaucoma , cancer , colorectal cancer, prostate cancer, renal cell cancer, dry eye disease , and macular degeneration . The compounds basal cell cancer, breast cancer, bone cancer, brain cancer, 10 can also be used to treat eye conditions that involve , are lymphoma, leukemia , melanoma, myeloma and other hema caused by, are effected by , or are exacerbated by MMP - 9 . tological cancers , and the like . The cancer can be primary , The compounds described herein can be used to treat metastatic , or both . The treatment of cancer using a com - inflammation , wherein the inflammation involves connec pound of the invention can affect (i . e ., inhibit or promote ) tive tissue , airway tissue , or central nervous system tissue. angiogenesis . The cardiovascular disease can be stroke , 15 The inflammation can be acute asthma , chronic asthma, aneurysm , ischemia or reperfusion injury . allergic asthma, or chronic obstructive pulmonary disease . A compound of a formula described herein , or a pharma - In one embodiment, the inflammation is arthritis . ceutically acceptable salt thereof, can be administered to a The compounds described herein can be used to treat an mammal ( e . g . , human ) alone or in conjunction with a second ophthalmologic disease . The term “ ophthalmologic disease ” agent, such as a neurological agent, or a pharmaceutically 20 or “ ophthalmologic disorder” refers to a disease or disorder acceptable salt thereof. Accordingly , the compound can be involving the anatomy and / or function of the visual system , administered in conjunction with a thrombolytic agent , such including but not limited to , glaucoma, retinal artery occlu as tPA to treat a disorder , disease , or condition as described sion , ischemic optic neuropathy and wet or dry macular herein . degeneration . The term " neurological agent” refers to a compound , 25 A neurological disorder can be an affective disorder ( e . g . , including chemical and biological compounds ( e . g . , pep - depression or anxiety ) . The term " affective disorder ” or tides , oligonucleotides and antibodies ) , that has an effect on " mood disorder” refers to a variety of conditions character the nervous system , e . g ., compounds capable of treating , ized by a disturbance in mood as the main feature . inhibiting or preventing disorders affecting the nervous The term “ depression ” refers to an abnormal mood dis system or compounds capable of eliciting a neurological 30 turbance characterized by feelings of sadness , despair, and and / or an ophthalmological disorder or symptoms thereof. discouragement. Depression refers to an abnormal emo Various studies indicate that MMP - 9 and MMP - 2 con - tional state characterized by exaggerated feelings of sadness , tribute in the disease process of stroke . Gelatinase inhibitors melancholy , dejection , worthlessness, emptiness , and hope can protect the neurovascular integrity of the brain from lessness that are inappropriate and out of proportion to ischemia or exogenous tPA thrombolysis by blocking deg - 35 reality . See , Mosby ' s Medical, Nursing & Allied Health radation of the basal membrane laminin and exerting anti - Dictionary, 5th Edition ( 1998 ) . Depression can be at least apoptotic effects on neurons. The selectivity of the com one of a major depressive disorder (single episode , recur pounds described herein therefore allow for the treatment of rent, mild , moderate , severe without psychotic features , both ischemic and hemorrhagic stroke . Additionally , com - severe with psychotic features, chronic , with catatonic fea bined treatment with selective gelatinase inhibitors and tPA 40 tures , with melancholic features, with atypical features , with can minimize neurotoxicity and hemorrhagic transformation postpartum onset , in partial remission , in full remission ) , associated with tPA use , thereby extending the window of dysthymic disorder, adjustment disorder with depressed treatment for reperfusion therapy of tPA . mood , adjustment disorder with mixed anxiety and For this approach to effectively treat stroke patients , it depressed mood , premenstrual dysphoric disorder, minor may require delivery of the gelatinase inhibitors by intra - 45 depressive disorder , recurrent brief depressive disorder, venous administration . Water - soluble prodrugs of second post- psychotic depressive disorder of schizophrenia , a major generation gelatinase inhibitors have been prepared as depressive disorder associated with Parkinson ' s disease , and described herein , and are amenable to intravenous admin - a major depressive disorder associated with dementia . istration . This novel therapeutic strategy by itself or in The term “ anxiety disorders ” refers to an excessive or combination with tPA can reduce injury and extend the time 50 inappropriate aroused state characterized by feelings of window for thrombolytic therapy in patients with stroke . apprehension , uncertainty , or fear. Anxiety disorders have The neurological disease can be one that arises from at been classified according to the severity and duration of their least one of painful neuropathy, neuropathic pain , diabetic symptoms and specific behavioral characteristics . Catego neuropathy , drug dependence , drug withdrawal, depression , ries include : Generalized anxiety disorder (GAD ) , which is anxiety , movement disorders , tardive dyskinesia , cerebral 55 long - lasting and low - grade; Panic disorder , which has more infections that disrupt the blood -brain barrier, meningitis , dramatic symptoms; Phobias ; Obsessive - compulsive disor stroke , hypoglycemia , cardiac arrest , spinal cord trauma, der (OCD ) ; Post - traumatic stress disorder (PTSD ); and head trauma , and perinatal hypoxia . The neurological dis - Separation anxiety disorder. ease can also be a neurodegenerative disorder. The neuro The neurological disorder can be pain associated depres logical disease can be epilepsy, Alzheimer ' s disease , Hun - 60 sion (PAD ) . The term “ pain associated depression ” refers to tington ' s disease, Parkinson ' s disease , multiple sclerosis , or a depressive disorder characterized by the co -morbidity of amyotrophic lateral sclerosis , as well as Alexander disease , pain and atypical depression . Specifically, the pain can be Alper 's disease , Ataxia telangiectasia , Batten disease (also chronic pain , neuropathic pain , or a combination thereof. known as Spielmeyer - Vogt - Sjogren -Batten disease ) , Cana - Specifically, the pain associated depression can include van disease , Cockayne syndrome, Corticobasal degenera - 65 atypical depression and chronic pain wherein the chronic tion , Creutzfeldt- Jakob disease , Kennedy ' s disease, Krabbe pain precedes the atypical depression . Alternatively , the pain disease , lewy body dementia , Machado - Joseph disease (Spi - associated depression can include atypical depression and US 9 ,951 , 035 B2 35 36 chronic pain wherein the atypical depression precedes the “ Neuropathic pain ” refers to pain associated with inflam chronic pain . The pain associated depression can include mation or degeneration of the peripheral nerves, cranial atypical depression and neuropathic pain . nerves , spinal nerves , or a combination thereof. The pain is “ Chronic pain ” refers to pain that continues or recurs over typically sharp , stinging , or stabbing . The underlying disor a prolonged period of time ( i. e ., > 3 mos. ), caused by various 5 der can result in the destruction of peripheral nerve tissue diseases or abnormal conditions, such as rheumatoid arthri - and can be accompanied by changes in the skin color, tis . Chronic pain may be less intense than acute pain . The temperature , and edema. See ,Mosby 's Medical, Nursing & person with chronic pain does not usually display increased Allied Health Dictionary, 5th Edition ( 1998 ) ; and Stedman 's pulse and rapid perspiration because the automatic reactions Medical Dictionary, 25th Edition (1990 ) . to pain cannot be sustained for long periods of time. Others The term “ diabetic neuropathy ” refers to a peripheral with chronic pain may withdraw from the environment and nerve disorder/ nerve damage caused by diabetes, including concentrate solely on their affliction , totally ignoring their peripheral , autonomic , and cranial nerve disorders/ damage family , their friends, and external stimuli. See, Mosby ' s associated with diabetes. Diabetic neuropathy refers to a Medical, Nursing & Allied Health Dictionary, 5th Edition 15 common complication of diabetes mellitus in which nerves ( 1998 ) . are damaged as a result of hyperglycemia ( high blood sugar “ Atypical depression ” refers to a depressed affect, with levels ). the ability to feel better temporarily in response to positive The term “ tardive dyskinesia ” refers to a serious, irre life effect (mood reactivity ), plus two or more neurovegeta - versible neurological disorder that can appear at any age . tive symptoms selected from the group of hypersomnia , 20 Tardive Dyskinesia , e. g ., Tourette 's syndrome, can be a side increased appetite or weight gain , leaden paralysis , and a effect of long - term use of antipsychotic /neuroleptic drugs . long standing pattern of extreme sensitivity to perceived Symptoms involve uncontrollable movement of various interpersonal rejection ; wherein the neurovegetative symp- body parts , including the body trunk , legs, arms, fingers , toms are present for more than about two weeks . It is mouth , lips, or tongue . appreciated that those of skill in the art recognize that the 25 The term " movement disorder ” refers to a group of neurovegatative symptoms can be reversed compared to neurological disorders that involve the motor and movement those found in other depressive disorders ( e . g ., melancholic systems, including, but are not limited to , Ataxia , Parkin depression ) ; hence the term “ atypical. ” son 's disease, Blepharospasm , Angelman Syndrome, Ataxia The term “ acute neurological disorder ” refers to a neu Telangiectasia , Dysphonia , Dystonic disorders , Gait disor rological disorder, as defined above , wherein the disorder 30 ders , Torticollis , Writer ' s Cramp , Progressive Supranuclear has a rapid onset which is followed by a short but severe Palsy , Huntington ' s Chorea , Wilson ' s Disease , Myoclonus , course , including , but not limited to , Febrile Seizures, Guil Spasticity , Tardive dyskinesia , Tics and Tourette syndrome lain - Barre syndrome, stroke , and intracerebral hemorrhag - and Tremors . ing (ICH ) . The term " cerebral infections that disrupt the blood -brain The term " chronic neurological disorder” refers to a 35 barrier ” refers to infections of the brain or cerebrum that neurological disorder , as defined above , wherein the disor - result in an alteration in the effectiveness of the blood -brain der lasts for a long period of time ( e . g . , more than about 2 barrier, either increasing or decreasing its ability to prevent, weeks ; specifically, the chronic neurological disorder can for example, substances and /or organisms from passing out continue or recur for more than about 4 weeks , more than of the bloodstream and into the CNS . about 8 weeks, or more than about 12 weeks ) or is marked 40 The term “ the blood - brain barrier” refers to a semi by frequent recurrence , including , but not limited to , narco - permeable cell layer of endothelial cells ( interior walls ) lepsy , chronic inflammatory demyelinating polyneuropathy, within capillaries of the central nervous system ( CNS) . The Cerebral palsy (CP ) , epilepsy, multiple sclerosis , dyslexia , blood - brain barrier prevents large molecules , immune cells , Alzheimer ' s disease and Parkinson ' s Disease . many potentially damaging substances, and foreign organ The term " trauma ” refers to any injury or shock to the 45 isms ( e . g . , viruses ) , from passing out of the bloodstream and body , as from violence or an accident . The term trauma also into the CNS (Brain and Spinal Cord ) . A dysfunction in the refers to any emotional wound or shock ,many of which may Blood - Brain Barrier may underlie in part the disease process create substantial, lasting damage to the psychological in MS (multiple sclerosis ). development of a person , often leading to neurosis . The term " meningitis ” refers to inflammation of the The term “ ischemic conditions ” refers to any condition 50 meninges of the brain and the spinal cord , most often caused which results in a decrease in the blood supply to a bodily by a bacterial or viral infection and characterized by fever, organ , tissue, or part caused by constriction or obstruction of vomiting , intense headache, and stiff neck . The term the blood vessels , often resulting in a reduction of oxygen to “ meningoencephalitis ” refers to inflammation of both the the organ , tissue, or part. The term “ hypoxic conditions” brain and meninges . refers to conditions in which the amount/ concentration of 55 The term “ stroke” refers to a sudden loss of brain function oxygen in the air , blood or tissue is low ( subnormal) . caused by a blockage or rupture of a blood vessel to the brain The term “ painful neuropathy ” or “ neuropathy ” refers to (resulting in the lack of oxygen to the brain ), characterized chronic pain that results from damage to or pathological by loss of muscular control, diminution or loss of sensation changes of the peripheral or central nervous system . Periph - or consciousness, dizziness , slurred speech , or other symp eral neuropathic pain is also referred to as painful neuropa - 60 toms that vary with the extent and severity of the damage to thy, nerve pain , sensory peripheral neuropathy, or peripheral the brain , also called cerebral accident , or cerebrovascular neuritis . With neuropathy, the pain is not a symptom of accident. The term " cerebral ischemia ” (or " stroke ” ) also injury, but rather the pain is itself the disease process . refers to a deficiency in blood supply to the brain , often Neuropathy is not associated with the healing process. resulting in a lack of oxygen to the brain . Rather than communicating that there is an injury some- 65 The term " cardiac arrest ” refers to a sudden cessation of where , the nerves themselves malfunction and become the heartbeat and cardiac function , resulting in a temporary or cause of pain . permanent loss of effective circulation . US 9 , 951 ,035 B2 37 38 The term " spinal cord trauma” refers to damage to the quite severely . Upper motor neuron variants ( e . g ., primary spinal cord that results from direct injury to the spinal cord lateral sclerosis ) are also included . itself or indirectly by damage to the bones and soft tissues The term “ glaucoma” refers to any of a group of eye and vessels surrounding the spinal cord . It is also called diseases characterized by abnormally high intraocular fluid Spinal cord compression ; Spinal cord injury ; or Compres - 5 pressure , damaged optic disk , hardening of the eyeball, and partial to complete loss of vision . The retinal ganglion cells sion of spinal cord . are lost in glaucoma. Some variants of glaucoma have The term " head trauma” refers to a head injury of the normal intraocular pressure (known also as low tension scalp , skull , or brain . These injuries can range from a minor glaucoma ). bump on the skull to a devastating brain injury . Head trauma| 10 The term “ retinal ischemia ” refers to a decrease in the can be classified as either closed or penetrating . In a closed blood supply to the retina . head injury , the head sustains a blunt force by striking The term “ ischemic optic neuropathy ” refers to a condi against an object . A concussion is a type of closed head tion that usually presents with sudden onset of unilaterally injury that involves the brain . In a penetrating head injury, reduced vision . The condition is the result of decreased an object breaks through the skull and enters the brainbrain .. 15 blood flow to the optic nerve ( ischemia ) . There are two basic The term “ perinatal hypoxia ” refers to a lack of oxygen types : arteritic and non - arteritic ischemic optic neuropathy. during the perinatal period (defined as the period of time Non -arteritic ischemic optic neuropathy is generally the occurring shortly before and after birth , variously defined as result of cardiovascular disease . Those patients at greatest beginning with completion of the twentieth to twenty eighth risk have a history of high blood pressure , elevated choles week of gestation and ending 7 to 28 days after birth ) . 20 terol, smoking , diabetes, or combinations of these. Arteritic The term “ hypoglycemic neuronal damage ” refers to ischemic optic neuropathy is a condition caused by the neuronal damage, for example , nerve damage, as a result of inflammation of vessels supplying blood to the optic nerve , a hypoglycemic condition (an abnormally low level of known as temporal arteritis . This condition usually presents glucose in the blood ). with sudden and severe vision loss in one eye , pain in the The term " epilepsy ” refers to any of various neurological 25 jaw with chewing , tenderness in the temple area , loss of disorders characterized by sudden recurring attacks of appetite , and a generalized feeling of fatigue or illness . motor, sensory , or psychic malfunction with or without loss The term “ macular degeneration ” refers to the physical of consciousness or convulsive seizures . disturbance of the center of the retina called the macula . The The term “ Alzheimer' s disease ” refers to a disease macula is the part of the retina which is capable of ourmost marked by the loss of cognitive ability, generally over a 30 acute and detailed vision .Macular degeneration is the lead period of 10 to 15 years , and associated with the develop i ng cause of legal blindness in people over age 55 (legal ment of abnormal tissues and protein deposits in the cerebral blindness means that a person can see 20 / 200 or less with cortex (known as plaques and tangles ) . eyeglasses .) Even with a loss of central vision , however, The term “ Huntington ' s disease” refers to a disease that is color vision and peripheral vision may remain clear. Vision hereditary in nature and develops in adulthood and ends in 35 loss usually occurs gradually and typically affects both eyes dementia . More specifically, Huntington ' s disease ( HD ) at different rates. results from genetically programmed degeneration of brain As used herein a “ demyelinating disorder ” refers to a cells , called neurons, in certain areas of the brain caused by medical condition where the myelin sheath is damaged . The a polyglutamine repeat in the DNA sequence of the gene myelin sheath surrounds nerves and is responsible for the encoding the protein huntingtin . This degeneration causes 40 transmission of impulses to the brain . Damage to the myelin uncontrolled movements , loss of intellectual faculties , and sheath may result in muscle weakness , poor coordination emotional disturbance . and possible paralysis . Examples of demyelinating disorders The term “ Parkinsonism ” refers to a disorder similar to include Multiple Sclerosis (MS ) , optic neuritis , transverse Parkinson ' s disease , but which is caused by the effects of a neuritis and Guillain - Barre Syndrome (GBS ) . In one medication , a different neurodegenerative disorder or 45 embodiment, when treating a demyelinating disorder , an another illness . The term " parkinsonism ” also refers to any MMP inhibitor is administered with an NMDAR antagonist condition that causes any combination of the types of ( e . g . , memantine ) or with B - interferon isoforms, copaxone movement abnormalities seen in Parkinson ' s disease by or Antegren ( natalizumab ) . Recently , it has been noted that damaging or destroying dopamine neurons in a certain area underlying neuronal damage can occur in demyelinating of the brain . 50 conditions such as MS , and therefore useful drugs may also The term “ amyotrophic lateral sclerosis ” (ALS ) , also protect the neurons instead or in addition to the myelin . called Lou Gehrig ' s disease and Motor Neuron Disease , The term “ multiple sclerosis ” refers to a chronic disease refers to a progressive , fatal neurological disease . The dis - of the central nervous system , which predominantly affects order belongs to a class of disorders known as motor neuron young adults . Viral and autoimmune etiologies are postu diseases . ALS occurs when specific nerve cells in the brain 55 lated . Genetic and environmental factors are known to and spinal cord that control voluntary movement gradually contribute to MS, but a specific cause for this disease is not degenerate (usually the “ upper” (in the cerebrocortex ) and yet identified . Pathologically , MS is characterized by the “ lower " ( in the spinal cord ) motor neurons, although some presence of areas of demyelination and T -cell predominant variants known as primary lateral sclerosis , apparently rep - perivascular inflammation in the brain white matter . Some resenting a separate disease , affect only the upper motor 60 axons may be spared from these pathological processes . The neurons ) . The loss of these motor neurons causes the disease begins most commonly with acute or subacute onset muscles under their control to weaken and waste away, of neurologic abnormalities. Initial and subsequent symp leading to paralysis . ALS manifests itself in different ways , toms may dramatically vary in their expression and severity depending on which muscles weaken first . Symptoms may over the course of the disease that usually lasts for many include tripping and falling , loss of motor control in hands 65 years . Early symptomsmay include numbness and / or par and arms, difficulty speaking , swallowing and /or breathing , esthesia , mono - or paraparesis , double vision , optic neuritis , persistent fatigue, and twitching and cramping, sometimes ataxia , and bladder control problems. Subsequent symptoms US 9 ,951 , 035 B2 39 40 also include more prominent upper motor neuron signs, i .e ., acceptable salt thereof, wherein the gelatinase inhibitor increased spasticity , increasing para - or quadriparesis . Ver accelerates the healing process of the skin wound . tigo , incoordination and other cerebellar problems, depres The invention also provides methods of inhibiting the sion , emotional lability , abnormalities in gait, dysarthria , progression of a skin wound associated disease state char fatigue and pain are also commonly seen . acterized by elevated levels of matrix metalloproteinases . The term “ sequelae of hyperhomocystinemia ” refers to a The methods can include administering to a mammal condition following as a consequence hyperhomocystine afflicted with a skin wound an effective amount of a gelati mia , meaning elevated levels of homocysteine . nase inhibitor, or a pharmaceutically acceptable salt thereof, The term " brain edema” refers to an excessive accumu effective to inhibit the progression of the skin wound in the lation of fluid in , on , around and /or in relation to the brain . 10 mammal. The term “ AIDS induced dementia ” or “ HIV - associated The invention further provides a method for enhancing dementia ” refers to dementia (deterioration of intellectual the rate of repair of a diabetic skin wound . Themethod can faculties , such as memory , concentration , and judgment , include administering to the skin wound an effective amount resulting from an organic disease or a disorder of the brain ) of a gelatinase inhibitor, or a pharmaceutically acceptable induced by AIDS (Acquired Immunodeficiency Syn - 15 salt thereof, wherein the rate of repair of the skin wound is dromean epidemic disease caused by an infection by enhanced , for example , compared to the rate of repair of a human immunodeficiency virus (HIV -1 , HIV -2 ) , a retrovirus skin wound not receiving administration of the gelatinase that causes immune system failure and debilitation and is inhibitor. often accompanied by infections such as tuberculosis ) . The invention additionally provides a dressing or patch The term “ HIV - related neuropathy ” refers to a neuropathy 20 for a chronic skin wound . The dressing or patch can include in a mammal infected with HIV were the neuropathy is an effective amount of a gelatinase inhibitor, or a pharma caused by infections such as CMV or other viruses of the ceutically acceptable salt thereof, and a pharmaceutically herpes family . Neuropathy is the name given to a group of acceptable carrier , diluent, or excipient. For example , the disorders whose symptoms may range from a tingling sen active can be included in an ointment base , where the sation or numbness in the toes and fingers to paralysis . 25 gelatinase inhibitor and the ointment base are combined and Neuropathy might more accurately be called “ neuropathies incorporated into a dressing . The dressing can a woven or because there are several types and can be painful. non -woven fabric and can further include a backing and / or The term “ retinopathy ” refers to any pathological disorder an adhesive . of the retina . In some embodiments , the effective amount of an MMP The term “ cognitive disorder ” refers to any cognitive 30 inhibitor described herein can be , for example , about 0 .01 to dysfunction , for example , disturbance of memory ( e . g ., about 50 mg per day , about 0 . 1 to about 10 mg per day, about amnesia ) or learning . 0 . 5 to about 5 mg per day, or about 0 . 5 to about 2 . 5 mg per The term “ neuronal injury associated with HIV infection ” day. The effective amount of the gelatinase inhibitor can be refers to damage /injury of nerve cells caused either directly applied , for example , topically , optionally in combination or indirectly by infection with HIV . 35 with other actives and / or carriers . The amount per day can The term “ dysfunction in cognition , movement and sen be an amount in a composition applied , for example , topi sation ” refers to abnormal or impaired functioning in cog - cally or transdermally , or it can be an amount administered nition (mental process of knowing , including aspects such as by another means, such as subdermally . For topical admin awareness , perception , reasoning, and judgment ), movement istration , the amount can also be about 0 . 01 to about 50 mg or sensation . 40 per day, about 0 . 1 to about 10 mg per day , about 0 . 5 to about Any of the above diseases, disorders, or conditions can be 5 mg per day , or about 0 . 5 to about 2 . 5 mg per 100 cm ? of treated by administering a compound or composition that wound on the surface of the patient being treated . includes a compound described herein to treat the disease , In some embodiments , the skin wound is a chronic skin disorder , or condition by selectively inhibiting matrix a wound . Subjects having wounds treatable by the methods metalloproteinase . 45 described herein include mammals , such as humans . In some Wound Therapy cases, the mammal can be suffering from diabetes , and the Selective matrix metalloproteinase (MMP ) inhibitors skin wound can be a chronic diabetic skin wound . The have been found to facilitate healing of wounds, including inhibitor can be delivered to the skin wound in a variety of diabetic wounds and chronic wounds. It has been discovered forms, such as in an ointment, or the administration of the that a number of selective inhibitor compounds significantly 50 inhibitor can be intraperitoneal, such as intravenous admin accelerate the healing process of various chronic wounds. istration . The evaluations described herein demonstrate that these The invention therefore provides therapeutic methods of compounds are indeed efficacious in accelerating the healing treating skin wounds in a mammal. The methods can include process in diabetic mammals. Notably , the therapy was administering to a mammal having a wound , such as a effective in diabetic mice but not in non - diabetic mice. The 55 chronic skin wound , an effective amount of a compound or non - diabetic mice treated with an MMP inhibitor failed to composition described herein . The invention also provides show any acceleration effect for their wound healing. These compounds useful for treating wounds of the integument compounds are the first discovered for this type of therapy . ( e . g . , skin ulcers and any break or damage to the integument ) There are no current clinical agents that can accelerate the or wounds as a result of surgery , which can include systemic wound healing process in diabetics , therefore the com - 60 treatment to aid the healing of such internal wounds . pounds, compositions , and methods described herein will be Marked upregulation of MMP - 2 and MMP - 9 is found in of significant importance to patients and practitioners in chronic wounds. Higher levels of MMP - 9 in chronic wound need of therapeutic methods for treating chronic wounds . fluid correlate with clinically more severe wounds . Reduced The invention thus provides methods of accelerating the levels of TIMP are also found in chronic wounds . As healing process of a skin wound . The methods can include 65 described herein , it has now been determined that selective administering to a mammal afflicted with a skin wound an gelatinase inhibitors can be effective in the treatment of effective amount of an MMP inhibitor, or a pharmaceutically chronic wounds . US 9 ,951 , 035 B2 41 42 The compositions and methods described herein can be or steroids . Chronic skin wounds can also be cause by used for aiding wound management. The term " wound bacterial, viral or fungal infections , or the presence of management” refers to therapeutic methods that induce foreign objects . and / or promote repair of a wound including , but not limited The term " diabetes ” refers to any of several metabolic to , arresting tissue damage such as necrotization , promoting 5 conditions characterized by the excessive excretion of urine tissue growth and repair , reduction or elimination of an and persistent thirst. The excess of urine can be caused by a established microbial infection of the wound and prevention deficiency of antidiuretic hormone , as in diabetes insipidus , of new or additionalmicrobial infection or colonization . The or it can be the polyuria resulting from the hyperglycemia term can further include reducing or eliminating the sensa that occurs in diabetes mellitus . tion of pain attributable to a wound . 10 The phrase " type 1 diabetes mellitus ” refers to the first of The therapeutic compositions for use in methods of the two major types of diabetes mellitus , characterized by wound management can include a surfactant that can useful abrupt onset of symptoms ( often in early adolescence ) , in cleaning a wound or contributing to bactericidal activity insulinopenia , and dependence on exogenous insulin . It of the administered compositions. Suitable surfactants results from a lack of insulin production by the pancreatic include, but are not limited to , phospholipids such as leci - 15 beta cells . With inadequate control, hyperglycemia , protein thin , including soy lecithin and detergents . The surfactant wasting , and ketone body production occur. The hypergly selected for application to a wound or skin surface will cemia leads to overflow glycosuria , osmotic diuresis , hyper typically be mild and will not lead to extensive irritation or osmolarity , dehydration , and diabetic ketoacidosis , which promote further tissue damage to the patient. can progress to nausea and vomiting , stupor, and potentially A " wound ” refers to an injury to the body , including but 20 fatal hyperosmolar coma. The associated angiopathy of not limited to an injury from trauma, violence , accident, or blood vessels ( particularly microangiopathy ) affects the reti surgery . A wound may occur due to laceration or breaking of nas, kidneys , and arteriolar basement membranes. Polyuria , a membrane ( such as the skin ) and usually damage to polydipsia , polyphagia , weight loss , paresthesias, blurred underlying tissues . A wound may occur in a topical location vision , and irritability can also occur. or internally . Chronic wounds may be caused by diseases , 25 The phrase " type 2 diabetes mellitus ” refers to the second including but not limited to diabetes ; diseases of internal of the two major types of diabetes mellitus , peaking in onset organs , including but not limited to diseases of the liver , between 50 and 60 years of age, characterized by gradual kidneys or lungs ; cancer, or any other condition that slows onset with few symptoms of metabolic disturbance ( glyco the healing process . suria and its consequences ) and control by diet, with or Natural healing occurs in clearly defined stages . Skin 30 without oral hypoglycemics but without exogenous insulin wounds of acute nature may heal in 1 - 3 weeks in a biological required . Basal insulin secretion is maintained at normal or process that restores the integrity and function of the skin reduced levels , but insulin release in response to a glucose and the underlying tissue. Such wounds may be the result of load is delayed or reduced . Defective glucose receptors on a scrape , abrasion , cut, graze, incision , tear , or bruise to the the pancreatic beta cells may be involved . It is often accom skin . If a wound does not heal in 4 - 12 weeks , it may be 35 panied by disease of blood vessels , particularly the large considered chronic . In the chase of chronic wounds, the ones , leading to premature atherosclerosis with myocardial wound may be attenuated at one of the stages of healing or infarction or stroke syndrome. fail to progress through the normal stages of healing . A Patients suffering from diabetes can develop chronic chronic wound may have been present for a brief period of wounds of the skin , internal wounds from surgery , or other time, such as a month , or it may have been present for 40 medical conditions that are not able to fully heal without the several years. aid of the treatments methods described herein . The phrase " chronic skin wound ” includes , but is not Combination Therapy limited to , skin ulcers, bed sores, pressure sores, diabetic In the following description , component “ ( b ) ” is to be ulcers and sores , and other skin disorders . Chronic skin understood to represent one or more agents as described wounds can be any size , shape or depth , and may appear 45 herein ( e . g . , a compound of Formula I ) . Thus , if components discolored as compared to normal , healthy skin pigment. ( a ) and ( b ) are to be treated the same or independently , each Chronic skin wounds can bleed , swell , seep pus or purulent agent of component ( b ) may also be treated the same or discharge or other fluid , cause pain or cause movement of independently . Components ( a ) and ( b ) may be formulated the affected area to be difficult or painful. Chronic skin together, in a single dosage unit (that is , combined together , wounds can become infected , producing elevated body 50 e . g . , in one lotion , cream , gel, ointment, or formulation for temperatures, as well as pus or discharge that is milky , injection ) as a combination product. When component ( a ) yellow , green , or brown in color , and is odorless or has a and ( b ) are not formulated together in a single dosage unit , pungent odor. If infected , chronic skin wounds may be red , the component ( a ) may be administered at the same time as tender, or warm to the touch . component (b ), or in any order. For example component ( a ) Chronic skin wounds can be caused by diabetes , poor 55 may be administered first, followed by administration of blood supply , low blood oxygen , by conditions where blood component ( b ) , or they may be administered in the reverse flow is decreased due to low blood pressure , or by conditions order. If component ( b ) contains more than one agent, e . g . , characterized by occluded , blocked or narrowed blood ves a thrombolytic agent and NSAID , these agents may be sels. A low oxygen supply can be caused by certain blood , administered together or separately in any order . When not heart, and lung diseases , and / or by smoking cigarettes . 60 administered at the same time, the administration of com Chronic skin wounds can also be the result of repeated ponent ( a ) and ( b ) can occur less than about one day , or less trauma to the skin , such as swelling or increased pressure in than about ten hours apart , or about one hour apart in some the tissues , or constant pressure on the wound area . Chronic embodiments . skin wounds can be caused by a weakened or compromised As is appreciated by a medical practitioner skilled in the immune system . A weakened or compromised immune 65 art, the dosage of the combination therapy of the invention system can be caused by increasing age , radiation , poor may vary depending upon various factors such as the nutrition , and / or medications, such as anti - cancer medicines pharmacodynamic characteristics of the particular agent and US 9 ,951 ,035 B2 43 44 its mode of administration , the age, health and weight of the selective serotonin reuptake inhibitors ( SSRIs ) , monoamine recipient, the nature and extent of the symptoms, the kind of oxidase inhibitor, anti - infective agent, nucleoside reverse concurrent treatment, the frequency of treatment, and the transcriptase , a protease inhibitor, or a thrombolytic agent effect desired , as described above . The proper dosage of such as tPA . components ( a ) and ( b ) will be readily ascertainable by a 5 Specifically , the MMP inhibitor can optionally be co medical practitioner skilled in the art . By way of general administered with at least one of the following : A beta guidance , typically a daily dosage may be about 10 milli - adrenergic blocking agent, carbonic anhydrase inhibitor, grams to about 1. 5 grams of each component. If component cholinesterase inhibitor, cholinergic (miotic ) , docosanoid , (b ) represents more than one compound, then typically a prostaglandin , tricyclic antidepressant, psychotherapeutic daily dosage may be about 10 milligrams to about 1 . 5 grams 10 agent, antianxiety agent, analgesic , anti- seizure agent, tri of each agent of component ( b ) . By way of general guidance , cyclic antidepressants having analgesic effect in neuropathic when the compounds of component ( a ) and component ( b ) pain , linolenic acid , coenzyme, vitamin , immunosuppressive are administered in combination , the dosage amount of each antimetabolite , antiviral, copolymer, barbiturate , benzodiaz component may be reduced by about 50 -80 % relative to the epine , GABA inhibitor, hydantoin , tranquilizer, anti- psy usual dosage of the component when it is administered alone 15 chotic , norephedrine , peptide, antibacterial, tissue plasmi as a single agent for the treatment of a disorder, and related nogen activator ( tPA ) , blood thinner /anticoagulant , symptoms, in view of synergistic effect of the combination . cardiostimulant, carbonic anhydrase inhibitor, ketoderiva Pharmaceutical kits useful for the treatment of disorders tive of carbamazepine, acetylcholinesterase , antipsychotic , described herein , and related symptoms, which include a alkaloid , GABA - B receptor agonist , benzodiazepine, anti therapeutically effective amount of a pharmaceutical com - 20 parkinsonian , antidepressant, CNS stimulant, receptor position that includes a compound of component ( a ) and one antagonist, beta adrenergic blocking agent, ergot derivatives or more compounds of component (b ) , in one or more sterile (anti migraine) , anticonvulsant, serotonin (5 -HT ) receptor containers , are also within the ambit of the invention . agonist , antimanic , SSRI, MAOI, aids adjunct anti - infective Sterilization of the container may be carried out using agent, antiviral, or protease inhibitor. conventional sterilization methodology well known to those 25 Additionally , the MMP inhibitor can optionally be co skilled in the art . Component ( a ) and component ( b ) may be administered with at least one of the following : in the same sterile container or in separate sterile containers . Timolol Maleate ; Timolol Hemihydrate ; Betaxolol HCl; The sterile containers of materials may include separate Metipranolol; Brimonidine Tartarate ; Brinzolamide; Dorzo containers , or one or more multi - part containers, as desired . lamide; Acetazolamide ; Echothiophate lodide; Pilocarpine Component ( a ) and component ( b ) , may be separate , or 30 HCl; Unoprostone Isopropyl ester; Latanoprost; Acampro physically combined into a single dosage form or unit as sate ; Amitriptyline ; Perphenazine ; Chlordiazepoxide ; Trim described above . Such kits may further include , if desired , ipramine Maleate ; Chlodiazepoxide HCl; Alprazolam ; one or more of various conventional pharmaceutical kit Hydroxyzine dihydrochloride; Meprobamate ; Doxipin HC1; components , such as for example , one or more pharmaceu Hydroxyzine Pamoate ; Aspirin ; Acetaminophen ; Ibuprofen ; tically acceptable carriers , additional vials for mixing the 35 Carbamazipine; Flupirtine ; Lamotrigine ; Phenytoin components, etc ., as will be readily apparent to those skilled Sodium ; Pentaxifylline; Thioctic Acid ; Levocarnitine ; Bio in the art. Instructions , either as inserts or as labels , indi - tin ; Nicotinic acid ; Taurine ; Verteporfin ; Azathioprine ; Inter cating quantities of the components to be administered , feron Beta 1 ; Interferon Beta 1 ; Cyclophosphamide ;Metho guidelines for administration , and/ or guidelines for mixing trexate; Neurmexane ; Mephobarbitol; Pentobarbitol ; the components , may also be included in the kit . 40 Lorazipam ; Clonazepam ; Chlorazeptate Dipotassium salt ; An MMP inhibitor described herein can optionally be Fosphenytoin Sodium ; Olanzapine ; Heloperidol; Trifluoper co -administered with a neuroprotectant drug , used , for izine ; Fluphenazine ; Phenylpropanol amine ; Pseudoephed example , in the treatment of Alzheimer ' s disease or other rine HCl; Imipramine ; Glucagon ; Glucagon -related peptide neurologic or ophthalmologic disorders (e .g ., glaucoma ), 1 ; Glucagon -related peptide - 2 ; Penicilin G , N , O , or V ; including , but not limited to , memantine or a derivative 45 Ampicillin , Chloramphenicol; Phorbol ; Heparin , D - glu thereof. cosamine with L - iduronic or D - glucuronic acids; Warfarin ; An MMP inhibitor described herein can optionally be Epinephrine ; Amiodarone ; Lidocaine ; Nitroglycerin , isosor co - administered with at least one of the followingowing :: bide dinitrate , amyl , butyl, isobutyl or various other nitrates An anti- glaucoma agent, beta adrenergic blocking agent, that have been shown to be neuroprotective ; Atenolol; carbonic anhydrase inhibitor, miotic agent, sympathomi- 50 Dexamethasone ; Prednisolone; Acetazolamide ; Phenytoin ; metic agent, acetylcholine blocking agent, antihistamine , Tiagabin HCl; Gabapentin ; Oxacarbazepine ; Tacrine; Done anti - viral agent, quinolone, anti - inflammatory agent, non - pezil ; Rivastigmine ; Heloperidol; Phenothiazine ; Reserpine ; steroidal anti- inflammatory agent, steroidal anti - inflamma- Tetrabenazene ; Bromocryptine; Tiapride ; Baclofen ; Diaz tory agent, antidepressant ( e . g ., serotonin reuptake inhibi epam ; Trihexyphenidyl HCl; Amitrityline ; Amphetamines ; tors , SSRIs ) , psychotherapeutic agent, anti- anxiety agent, 55 Methylphenidate ; Amitriptylinec ; Clomipramine ; Dolas analgesic , antiseizure agent, anti - convulsant, gabapentine , etron ; Granisetron ; Huperzine ; Metoclopramide ; Prochlo anti - hypertensive agent, benzoporphyrin phtosensitiser, rperazine ; Dexamethasone ; Timolol Hydrogen maleate salt ; immunosuppressive antimetabolite , barbiturate , benzodiaz Propanolol; Isometheptine ; Atenolol; Metoprolol; Nadolol; epine, GABA inhibitors , hydantoin , anti -psychotic , neuro - Ergotamine ; Dihydroargotamine ; Naratriptan ; Sumatriptan ; laptic, antidysknetic , adrenergic agent, tricyclic antidepres - 60 Rizatriptan ; Zolmitriptan ; Imipramine HCl; Dopamine ; Clo sant , anti -hypoglycemic , glucose solution , plypeptide zapine ; Valproic Acid , Amitriptylinec ; Imipramine HCl; hormone , antibiotic , thrombolytic agent, blood thinner, anti - Imipramine Pamoate ; Clomipramine ; Amphetamine ; Meth arrhythmic agent, corticosteroid , seizure disorder agent, ylphenidate ; Phenytoin ; Phenobarbital; Amitryptyline ; Imi anticholinesterase , dopamine blocker, antiparkinsonian pramine Pamoate ; Nortrityline ; Trazodone; Nefazodone ; agent, muscle relaxant, anxiolytic muscle relaxant, CNS 65 Sertraline; Fluoxetine ; Paroxetine; Phenalzine ; Tranylcy stimulant, antiemetic , beta adrenergic blocking agents , ergot promine ; Erythropoietin , a glycoprotein ; Immunoglobulins derivative , isometheptene , antiserotonin agent, analgesic , ( gamma globulins ) ; Tetrahydrocannabinols ; Alitretinoin ; US 9 ,951 , 035 B2 45 46 Lamivudin ; Stavudin ; Zalcitabine ; Abacavir ; Ritonavir ; fonates , propanesulfonates, naphthalene - 1 - sulfonates , naph Indinavir ; and Nelfinavir ; the chemical names of which are thalene - 2 -sulfonates , and mandelates . well known in the art and are also described in U . S . If the compound of a formula described herein contains a Publication No. 2009/ 0209615 (Liption et al .) , which is basic nitrogen , the desired pharmaceutically acceptable salt incorporated herein by reference . The MMP inhibitor can be 5 may be prepared by any suitable method available in the art . administered with an additional MMP inhibitors, including for example , treatment of the free base with an inorganic a compounds disclosed in U . S . Pat. No . 6 ,703 ,415 (Mobash acid , such as hydrochloric acid , hydrobromic acid , sulfuric ery et al. ) and U .S . Pat. No . 7 , 928 , 127 (Lee et al. ); PCT Publication No. WO 2011 / 026107 (Mobashery et al . ) ; and acid , sulfamic acid , nitric acid , boric acid , phosphoric acid , U . S . Publication No . 2013 /0064878 (Chang et al. ) ; which 10 and the like , or with an organic acid , such as acetic acid , patent documents are incorporated herein by reference . Any phenylacetic acid , propionic acid , stearic acid , lactic acid , one or more of the above compounds can be used in a ascorbic acid , maleic acid , hydroxymaleic acid , isethionic pharmaceutically acceptable salt form , solvate form (e . g ., a acid , succinic acid , valeric acid , fumaric acid , malonic acid , mono - or di- hydrate ) , or any combination thereof. pyruvic acid , oxalic acid , glycolic acid , salicylic acid , oleic Pharmaceutical Salts and Solvates 15 acid , palmitic acid , lauric acid , a pyranosidyl acid , such as The invention also includes pharmaceutically acceptable glucuronic acid or galacturonic acid , an alpha- hydroxy acid , salts and /or solvates of the compounds represented by a such as mandelic acid , citric acid , or tartaric acid , an amino formula described herein , such as those described above and acid , such as aspartic acid or glutamic acid , an aromatic acid , of the specific compounds exemplified herein , and methodsthods 20 Sucsuch as benzoic acid , 2 - acetoxybenzoic acid , naphthoic acid , or cinnamic acid , a sulfonic acid , such as laurylsulfonic acid , of treatment using such salts and /or solvates. p -toluenesulfonic acid , methanesulfonic acid , ethanesulfo A “ pharmaceutically acceptable salt” is intended to mean nic acid , any compatible mixture of acids such as those a salt of a free acid or base of a compound represented by given as examples herein , and any other acid and mixture a formula described herein that is non - toxic , biologically 25 thereof that are regarded as equivalents or acceptable sub tolerable , or otherwise biologically suitable for administra stitutes in light of the ordinary level of skill in this technol tion to the subject . See , generally , S . M . Berge , et al ., ogy . “ Pharmaceutical Salts ” , J . Pharm . Sci. , 1977 , 66 : 1 - 19, and If the compound of a formula described herein includes an Handbook of Pharmaceutical Salts , Properties, Selection , acid moiety , such as a carboxylic acid or sulfonic acid , the and Use , Stahl and Wermuth , Eds. , Wiley - VCH and VHCA , 30 desired pharmaceutically acceptable salt may be prepared by Zurich , 2002 . Preferred pharmaceutically acceptable salts any suitable method , for example, treatment of the free acid are those that are pharmacologically effective and suitable with an inorganic or organic base , such as an amine (pri for contact with the tissues of patients without undue tox - mary , secondary or tertiary ), an alkali metal hydroxide , icity , irritation , or allergic response . alkaline earth metal hydroxide , any compatible mixture of Pharmaceutically acceptable salts include the addition bases such as those given as examples herein , and any other salts of inorganic acids such as hydrochloride , hydrobro - base and mixture thereof that are regarded as equivalents or mide , hydroiodide, halide, sulfate , phosphate , carbonate , acceptable substitutes in light of the ordinary level of skill in bicarbonate , diphosphate and nitrate or of organic acids such this technology . Illustrative examples of suitable salts as acetate , malonate , maleate, fumarate , tartrate , succinate , 10 include organic salts derived from amino acids, such as citrate , lactate , benzoate , ascorbate , tosylate , mesylate , tri - glycine and arginine , ammonia , carbonates, bicarbonates, flate , palmoate , stearate , a -ketoglutarate , and a - glycero - primary , secondary , and tertiary amines , and cyclic amines , phosphate . Also within the scope of the present invention , such as benzylamines, pyrrolidines , piperidine, morpholine , when they can be used , are the salts formed from bases such and piperazine , and inorganic salts derived from sodium , as sodium or potassium hydroxide . For other examples of 45 calcium , potassium , magnesium , manganese, iron , copper , pharmaceutically acceptable salts, reference can be made to zinc , aluminum , and lithium . “ Salt selection for basic drugs” , Int. J. Pharm . ( 1986 ), 33 , The term " solvate ” refers to a solid compound that has 201 - 217 . one or more solvent molecules associated with its solid A compound of a formula described herein may possess structure . Solvates can form when a compound is crystal a sufficiently acidic group , a sufficiently basic group , or both 50 lized from a solvent, wherein one or more solventmolecules types of functional groups, and accordingly react with a become integral part( s ) of the crystal. The compounds of a number of inorganic or organic bases, and inorganic and formula described herein can be solvates, for example , organic acids, to form a pharmaceutically acceptable salt . ethanol solvates . Likewise , a " hydrate ” refers to a solid Examples of pharmaceutically acceptable salts include sul compound that has one or more water molecules associated fates , pyrosulfates , bisulfates , sulfites , bisulfites, phosphates, with its solid structure . A hydrate is a subgroup of solvates . monohydrogen -phosphates , dihydrogenphosphates, meta Hydrates can form when a compound is crystallized from phosphates , pyrophosphates , chlorides , bromides , iodides , water, wherein one or more water molecules become inte acetates, behenates , besylates, propionates , decanoates , gral part ( s ) of the crystal. The compounds of a formula caprylates , acrylates , formates, isobutyrates , caproates, hep - an described herein can be hydrates . tanoates , propiolates, oxalates , malonates, succinates , sub - Pharmaceutical Formulations erates , sebacates, fumarates , maleates , butyne - 1, 4 -dioates , The compounds described herein can be used to prepare hexyne - 1 , 6 - dioates, benzoates, chlorobenzoates, methylben therapeutic pharmaceutical compositions, for example , by zoates, dinitrobenzoates, hydroxybenzoates, methoxybenzo combining the compounds with a pharmaceutically accept ates, phthalates, sulfonates, xylenesulfonates, phenylac - 65 able diluent , excipient, or carrier . The compounds may be etates, phenylpropionates , phenylbutyrates , citrates , lactates , added to a carrier in the form of a salt or solvate . For y -hydroxybutyrates , glycolates, tartrates , methane - sul example , in cases where compounds are sufficiently basic or US 9 ,951 , 035 B2 47 48 acidic to form stable nontoxic acid or base salts , adminis - tion , the active compound may be incorporated into sus tration of the compounds as salts may be appropriate . tained - release preparations and devices . Examples of pharmaceutically acceptable salts are organic The active compound may be administered intravenously acid addition salts formed with acids that form a physiologi- or intraperitoneally by infusion or injection . Solutions of the cal acceptable anion , for example , tosylate , methanesul- active compound or its salts can be prepared in water, fonate , acetate , citrate , malonate , tartrate , succinate , benzo optionally mixed with a nontoxic surfactant. Dispersions can ate , ascorbate , a -ketoglutarate , and (3 -glycerophosphate . be prepared in glycerol, liquid polyethylene glycols , triace Suitable inorganic salts may also be formed , including tin , or mixtures thereof, or in a pharmaceutically acceptable hydrochloride , halide, sulfate , nitrate , bicarbonate , and car - 10 oil . Under ordinary conditions of storage and use, prepara bonate salts . tions may contain a preservative to prevent the growth of Pharmaceutically acceptable salts may be obtained using microorganisms. standard procedures well known in the art , for example by Pharmaceutical dosage forms suitable for injection or reacting a sufficiently basic compound such as an amine with infusion can include sterile aqueous solutions, dispersions, a suitable acid to provide a physiologically acceptable ionic 15 or sterile powders comprising the active ingredient adapted compound . Alkali metal ( for example , sodium , potassium or for the extemporaneous preparation of sterile injectable or lithium ) or alkaline earth metal ( for example , calcium ) salts infusible solutions or dispersions, optionally encapsulated in of carboxylic acids can also be prepared by analogous liposomes . The ultimate dosage form should be sterile , fluid methods . 20 and stable under the conditions of manufacture and storage . The compounds of the formulas described herein can be The liquid carrier or vehicle can be a solvent or liquid formulated as pharmaceutical compositions and adminis - dispersion medium comprising , for example , water , ethanol , tered to a mammalian host, such as a human patient, in a a polyol ( for example , glycerol, propylene glycol, liquid variety of forms. The forms can be specifically adapted to a polyethylene glycols , and the like ), vegetable oils , nontoxic chosen route of administration , e . g ., oral or parenteral 25 glyceryl esters, and suitable mixtures thereof. The proper administration , by intravenous, intramuscular, topical or fluidity can be maintained , for example , by the formation of subcutaneous routes . liposomes, by the maintenance of the required particle size The compounds described herein may be systemically in the case of dispersions, or by the use of surfactants . The administered in combination with a pharmaceutically 20 prevention of the action of microorganisms can be brought acceptable vehicle , such as an inert diluent or an assimilable 30 edible carrier . For oral administration , compounds can be about by various antibacterial and / or antifungal agents , for enclosed in hard or soft shell gelatin capsules , compressed example , parabens, chlorobutanol , phenol, sorbic acid , into tablets, or incorporated directly into the food of a thimerosal, and the like. In many cases, it will be preferable patient' s diet. Compounds may also be combined with one to include isotonic agents , for example , sugars , buffers , or or more excipients and used in the form of ingestible tablets , 35 sodium chloride . Prolonged absorption of the injectable buccal tablets , troches , capsules , elixirs , suspensions , syr compositions can be brought about by agents delaying ups, wafers , and the like . Such compositions and prepara - absorption , for example , aluminum monostearate and / or tions typically contain at least 0 . 1 % of active compound . gelatin . The percentage of the compositions and preparations can 10 Sterile injectable solutions can be prepared by incorpo vary and may conveniently be from about 0 .5 % to about rating the active compound in the required amount in the 60 % , about 1 % to about 25 % , or about 2 % to about 10 % , of appropriate solvent with various other ingredients enumer the weight of a given unit dosage form . The amount of active ated above, as required, optionally followed by filter steril compound in such therapeutically useful compositions can ization . In the case of sterile powders for the preparation of be such that an effective dosage level can be obtained . 45 sterile injectable solutions , methods of preparation can The tablets , troches, pills , capsules , and the like may also include vacuum drying and freeze drying techniques, which contain one or more of the following : binders such as gum yield a powder of the active ingredient plus any additional tragacanth , acacia , corn starch or gelatin ; excipients such as desired ingredient present in the solution . dicalcium phosphate ; a disintegrating agent such as corn For topical administration , compounds may be applied in starch , potato starch , alginic acid and the like; and a lubri - 50 pure form , e . g ., when they are liquids. However , it will cant such as magnesium stearate . A sweetening agent such generally be desirable to administer the active agent to the as sucrose , fructose , lactose or aspartame; or a flavoring skin as a composition or formulation , for example , in agent such as peppermint, oil of wintergreen , or cherry combination with a dermatologically acceptable carrier, flavoring , may be added . When the unit dosage form is a which may be a solid , a liquid , a gel , or the like . capsule, it may contain , in addition to materials of the above » Useful solid carriers include finely divided solids such as type , a liquid carrier, such as a vegetable oil or a polyeth - talc , clay , microcrystalline cellulose , silica , alumina , and the ylene glycol. Various other materials may be present as like . Useful liquid carriers include water , dimethyl sulfoxide coatings or to otherwise modify the physical form of the (DMSO ) , alcohols , glycols, or water - alcohol/ glycol blends , solid unit dosage form . For instance , tablets , pills , or cap - in which a compound can be dissolved or dispersed at sules may be coated with gelatin , wax , shellac or sugar and effective levels , optionally with the aid of non - toxic surfac the like . A syrup or elixir may contain the active compound , tants . Adjuvants such as fragrances and additional antimi sucrose or fructose as a sweetening agent, methyl and propyl crobial agents can be added to optimize the properties for a parabens as preservatives , a dye and flavoring such as cherry given use. The resultant liquid compositions can be applied or orange flavor. Any material used in preparing any unit 65 from absorbent pads, used to impregnate bandages and other dosage form should be pharmaceutically acceptable and dressings , or sprayed onto the affected area using a pump substantially non - toxic in the amounts employed . In addi type or aerosol sprayer. US 9 ,951 ,035 B2 49 50 Thickeners such as synthetic polymers , fatty acids, fatty EXAMPLES acid salts and esters , fatty alcohols , modified celluloses, or modified mineral materials can also be employed with liquid ABBREVIATIONS. ADAM , a disintegrin and metallo carriers to form spreadable pastes, gels , ointments , soaps, proteinase ; AUC , area under the concentration - time curve , and the like , for application directly to the skin of the user. 3 BBB , blood -brain barrier ; Boc , t- butoxycarbonyl ; m -CPBA , Examples of dermatological compositions for delivering meta -chloroperbenzoic acid ; CNS , central nervous system ; active agents to the skin are known to the art ; for example , DIEA , N , N -diisopropylethylamine ; DMF, dimethylforma mide; DMSO , dimethyl sulfoxide; ESI , electrospray ioniza see U . S . Pat. No . 4 ,992 ,478 (Geria ) , U . S . Pat. No. 4 , 820 ,508 tion ; HCTU , O - ( 1H - 6 - Chlorobenzotriazole - 1 - yl) - 1 , 1 , 3 , 3 - te (Wortzman ), U .S . Pat. No. 4 ,608 , 392 ( Jacquet et al. ) , and 10 tramethyluronium hexafluorophosphate ; MMP, matrix U .S . Pat . No. 4 ,559 , 157 (Smith et al .) . Such dermatological metalloproteinase ; MOCAC, (7 -methoxycoumarin - 4 -yl ) compositions can be used in combinations with the com acetyl ; MRM , multiple reaction monitoring ; MS, mass spec pounds described herein where an ingredient of such com trometry ; TEA , triethylamine ; THF, tetrahydrofuran ; TIMP, positions can optionally be replaced by a compound tissue inhibitor of metalloproteinase ; TLC , thin - layer chro described herein , or a compound described herein can be 15 matography ; UPLC , ultra- performance liquid chromatogra added to the composition . phy. Useful dosages of the compounds described herein can be determined by comparing their in vitro activity , and in vivo Example 1 . Selective Water -Soluble and activity in animal models . Methods for the extrapolation of 30 Slow - Binding Matrix Metalloproteinase - 2 and - 9 effective dosages in mice , and other animals , to humans are Inhibitors that Cross the Blood - Brain Barrier known to the art ; for example , see U . S . Pat. No . 4 , 938 , 949 ( Borch et al. ) . The amount of a compound , or an active salt The invention provides selective MMP inhibitors such as or derivative thereof, required for use in treatment will vary p -aminomethyloxadiazol ( 4 ) , p - aminomethyl (5 , ND - 336 ) , not only with the particular compound or salt selected but 25 p -acetamidomethyl (6 , ND -378 ), and p - guanidino ( 7 ) ana also with the route of administration , the nature of the logs of compound SB - 3CT ( 1 ) , and methods for their use . condition being treated , and the age and condition of the The compounds are 10 - to 14 , 000 - fold more water -soluble patient, and will be ultimately at the discretion of an than 1 , retain slow -binding inhibition behavior toward attendant physician or clinician . MMP - 2 , and can cross the blood brain barrier (BBB ) . The The compound can be conveniently administered in a unit 30 p -acetamidomethyl analog ( compound 6 ) is a selective dosage form , for example , containing 5 to 1000 mg /m² , conveniently 10 to 750 mg/ m ” , most conveniently, 50 to 500 nanomolar slow -binding inhibitor of MMP- 2 , which does mg/ m ? ofactive ingredient per unit dosage form . The desired not inhibit the closely related MMP - 9 or MMP- 14 . Because dose may conveniently be presented in a single dose or as of the slow dissociation of compound 6 from the target divided doses administered at appropriate intervals , for 35 MMP- 2 , it results in sustained inhibition of MMP - 2 even example , as two , three , four or more sub - doses per day . The when concentrations of 6 fall below the K? value . This sub - dose itself may be further divided , e . g . , into a number of inhibitor is a useful tool in therapeutic intervention and in discrete loosely spaced administrations . investigations of the role of MMP - 2 in neurological dis The invention provides therapeutic methods of treating eases . The p -aminomethyl derivative (compound 5) is a cancer in a mammal, which involve administering to a 40 water- soluble nanomolar slow - binding inhibitor ofMMP - 2 , mammal having cancer an effective amount of a compound MMP - 9, and MMP - 14 and has residence times for inhibition or composition described herein . A mammal includes a of these enzymes 6 - to 7 - fold longer than those of the tissue primate , human , rodent, canine , feline, bovine , ovine , inhibitors of metalloproteinase 1 or 2 ( TIMP- 1 or TIMP - 2 ) equine , swine , caprine , bovine and the like . Cancer refers to bound to MMP - 9 . any various type ofmalignant neoplasm , for example , colon 45 Chemistry . cancer, breast cancer , melanoma and leukemia , and in gen - The syntheses of the p -aminomethyloxadiazol ( 4 ) , eral is characterized by an undesirable cellular proliferation , p -aminomethyl ( 5 , ND - 336 ), p -acetamidomethyl ( 6 , e. g. , unregulated growth , lack of differentiation , local tissue ND -378 ), and p -guanidino ( 7) derivatives of 1 are outlined invasion , and metastasis . The ability of a compound of the below in Scheme 1 . Allylation of 4 -mercaptophenol (8 ) gave invention to treat cancer may be determined by using assays 50 compound 9 ( Ikejiri et al. , J. Biol. Chem . 2005 , 280 , well known to the art. For example , the design of treatment 33992 - 34002) , which was allowed to react with 4 - fluo protocols , toxicity evaluation , data analysis , quantification robenzonitrile to afford the diphenyl ether 10 . Reaction of 10 of tumor cell kill, and the biological significance of the use of transplantable tumor screens are known . The invention with hydroxylamine , followed by the addition of Boc also provides therapeutic methods of treating wounds such 55 glycine to the amidoxime generated the oxadiazol derivative as chronic wounds and wounds of diabetic patients , and 12, which was further oxidized to the corresponding oxirane neurological conditions such as stroke and traumatic brain 13 and subsequently reacted with thiourea to yield the injury , as well as compounds useful for evaluating the role Boc -protected thiirane 14 . Boc - deprotection using 4 N HC1 of MMPs in various conditions and disorders , such as the in 1 ,4 -dioxane gave the desired thiirane 4 . Reduction of 10 conditions and disorders described herein . so with LiAlH4, followed by Boc -protection gave 15 . Com The following Examples are intended to illustrate the pound 5 was synthesized from 15 using similar procedures above invention and should not be construed as to narrow its used for the synthesis of 4 from 12 . N - acetylation of 5 using scope . One skilled in the art will readily recognize that the acetyl chloride, in the presence of TEA , afforded compound Examples suggest many other ways in which the invention 6 . Reaction of 2 with N , N - di- Boc - 1H -pyrazole - 1 - carbox could be practiced . It should be understood that numerous 65 amidine, using N ,N -diisopropylethylamine (DIEA ) as base , variations and modifications may be made while remaining readily provided thiirane 18 , which was then Boc- depro within the scope of the invention . tected with HCl gas to afford the p - guanidino derivative 7 . US 9, 951, 035 B2 51 52 Scheme 1 . Syntheses of the inhibitors 4 , 5 , 6 and 70 .

OH LOH b N = HS NC NH2 - ~ . - caroan10 11 h , i

HN Boc Boc Boc wory woran15 peran

??? ??? HN HN maloBoc guardanapo 7 13

53 , 6 Boca

2 - Boc N NH

18 a Reagents and conditions: (a ) allyl bromide, K2CO3, 94 % ; ( b ) 4 - fluorobenzonitrile , Cs2CO3, DMF, 100° C ., 82 % ; (c ) hydroxylamine (50 % in water ), EtOH , reflux ; ( d ) Boc - glycine, HCTU , DIEA , DMF , 100° C . , 78 % in two steps ; ( e ) m - CPBA , CH , Cl2 , 0° C . to room temperature (~ 22° C .) , 88 d ,, 36 - 74 % ;; ((f ) thiourea , MeOH /CH2Cl2 ,, room temperature , 24 h , 58 -78 % ; ( g ) 4N HCl ( in 1 , 4 -dioxane ) , ETOAc/ CH2Cl2 , 0° C . to room temperature , 24 h , 98 - 99 % ; (h ) LiAlH4, THF; (i ) (Boc ) 20 , 12, MeOH /CH2Cl2 , 55 % in two steps; (j ) acetyl chloride, TEA , THF, 0° C . to room temperature ( ~ 22° C .) , 66 % ; ( k ) N , N '- di - Boc -1H -pyrazole - 1 -carboxamidine , DIEA ( 2 . 2 eq ), THF, room temperature , 24 h , 68 % ; ( 1) HCl ( gas) , EtOAc/ CH2Cl2 , 0° C ., 3 min , 95 % .

Experimental Section 4 -( 4 -( Allylthio )phenoxy )benzonitrile (10 ) Chemistry. 45 A mixture of 9 (1 .45 g, 8 .72 mmol ), 4 - fluorobenzonitrile ( 1 .01 g , 8 . 38 mmol) , and Cs2CO3 ( 4 .26 g , 13 . 1 mmol) in All reactions were performed under nitrogen atmosphere , DMF ( 50 mL ) was heated at 100° C . for 3 . 5 h . After the unless noted otherwise . ' H and 13C NMR spectra were addition of saturated aqueous LiBr (250 mL ) , the mixture recorded on Varian INOVA - 500 (Varian Inc. , Palo Alto , was extracted with hexanes /EtoAc ( 9 : 1 ) . The combined Calif ., USA ), Varian UnityPlus 300 spectrometer (Varian 50 organic layers were washed with water and brine , dried over Inc ., Palo Alto , Calif ., USA ) , Bruker AVANCE III HD 500 anhydrous Na , SO2, and concentrated under reduced pres ( Bruker Corporation , Billerica , Mass ., USA ) , or Bruker sure. The resultant residue was purified by silica gel chro AVANCE III HD 400 (Bruker Corporation , Billerica , Mass . , matography ( hexanes/ EtOAc, 97 : 3 ) to give 5 ( 1 .84 g , 82 % ) USA ). TLC silica gel 60 F254 aluminum sheets (EMD 55 as an oil . ' H NMR ( 300 MHz, CDC12 ) 8 7 .77 - 7 .52 ( m , 2H ) , Millipore Corporation , Billerica, Mass ., USA ) were used for 7 .49 - 7 .30 ( m , 2H ) , 7 . 14 - 6 .82 ( m , 4H ) , 5 .87 (ddt , J = 16 . 9 , thin - layer chromatography. Flash chromatography was per 10 . 0 , 6 .9 Hz, 1H ) , 5 . 29 -4 . 93 (m , 2H ), 3 .53 (dt , J = 6 .9 , 1. 1 Hz, formed with an automated chromatograph system : Combi 2H ) . 13C NMR ( 126 MHz , CDCI, ) 8 161 . 6 , 153 . 9 , 134 . 4 , flash RF 200i UV /Vis ( Teledyne Isco , Lincoln , Nebr ., USA ) . 133 .7 , 132 . 7 , 132 .4 , 121. 0, 119. 0 , 118 . 2 , 118 . 1, 106 . 3 , 38 . 2 . High -resolution mass spectra were obtained by ESI ioniza - 60 HRMS (ESI + , m / z ) : calcd for C , H , NO M + H ] + . tion , using a BrukermicroTOF/ Q2 mass spectrometer 268 . 0791 : found 268 .0799 ( BrukerDaltonik , Bremen , Germany ) . Purity of the prepared compounds was in general > 95 % , as confirmed by UPLC . ( E ) - 4 - ( 4 - ( Allylthio )phenoxy ) - N '- hydroxybenzimid Conditions are detailed in the UPLC section . 4 - ( Allylthio ) amide ( 11) phenol ( 9 ) was prepared as previously described ( Ikejiri et 65 al. , J . Biol. Chem . 2005 , 280 , 33992 - 34002 ; Goux et al. , C . ; A solution of 10 ( 865 mg, 3 . 24 mmol) and hydroxylamine Tetrahedron 1994 , 50 , 10321 - 10330 ) . (793 ?L , 50 % in water , 12 . 9 mmol) in EtOH ( 40 mL ) was US 9 ,951 ,035 B2 53 54 refluxed for 1 . 5 h . The solvent was evaporated under was purified by silica gel chromatography ( 130 . 1 mg, 78 % ) . reduced pressure to give 11 as a white solid , which was used ' H NMR (300 MHz , CDC1z) 8 8 . 13 ( d , J = 8 . 9 Hz , 2H ) , 7 .92 directly in the next step without further purification . ( d , J = 8 .7 Hz , 2H ), 7 . 18 ( d , J= 8. 5 Hz, 4H ), 5. 30 ( s, b , 1H ), 4 .65 ( s , 2H ) , 3 . 52 (dd , J = 14 . 0 , 5 . 8 Hz, 1H ) , 3 .22 (dd , J = 14 . 3 , t - Butyl ( ( 3 - ( 4 - ( 4 - (allylthio )phenoxy ) phenyl ) - 1 , 2 , 4 - 5 7 . 6 Hz, 1H ) , 3 . 16 - 2 .99 ( m , 1H ) , 2 .56 (dd , J = 6 . 0 , 1 . 6 Hz, 1H ) , oxadiazol- 5 - yl) methyl ) carbamate (12 ) 2 . 18 (dd , J = 5 . 0 , 1 . 6 Hz, 1H ) , 1 . 48 ( s , 9H ) . 13C NMR ( 75 MHz, CDC13 ) 8 177 . 0, 167 .9 , 162 .1 , 157 .9 , 143. 2 , 133 . 2 , DIEA ( 1 .05 mL , 5 .93 mmol) was added to a solution of 131. 2 , 130 . 0 , 123 .5 , 120 .5 , 118 . 8 , 77 .5 , 62 .9 , 37 .5 , 28 . 5 , Boc - glycine (692 mg, 3 . 95 mmol ) and 11 ( 989 mg, 3 . 29 26 . 3 , 24 . 4 . HRMS (ESI + , m / z ) : calcd for C2 H2N2O . S , mmol) in DMF ( 16 mL ) . HCTU ( 1 .64 g , 3 . 95 mmol) was 10 IM + H1+ 504 . 1258 : found . 504 . 1247 . then added to the resulting mixture at room temperature and stirred at 100° C . for 3 h . At this time, the TLC showed ( 3 - (4 - (4 - (( Thiiran - 2 - ylmethyl) sulfonyl) phenoxy ) complete conversion of starting materials . The solution was partitioned between EtoAc and LiBr aqueous solution . The phenyl ) - 1 , 2 ,4 - oxadiazol- 5 - yl) methanamine . HCl salt aqueous solution was extracted with EtoAc and the com - 15 ( 4 ) bined organic layers were washed with saturated NaHCO , and brine , dried over anhydrous Na2SO4 and concentrated in HCl ( 0 . 7 mL , 4 N in 1 , 4 - dioxane ) was added to a solution vacuo . The residue was purified by silica gel chromatogra of thiirane 14 ( 71 mg, 0 . 14 mmol) in CH C12 / EtOAC ( 1 : 1 , 4 phy (hexanes / EtoAc, 85 : 15 ) to give 12 ( 1. 13 g , 78 % in two mL ) . After stirring at room temperature for 24 h , the mixture steps ). ' H NMR (500 MHz , CDC13 ) 8 8 .03 (d , J = 8 .6 Hz, 20 was" concentrated under reduced pressure. The resulting 2H ), 7 .43 - 7 . 35 ( m , 2H ), 7 .05 (d , J= 8 .7 Hz, 2H ), 7 .02 - 6 .97 crude compound was triturated with diethyl ether, and the ( m , 2H ) , 5 . 95 - 5 . 80 ( m . 1H ) . 5 . 22 ( s . b . 11 ) . 5 . 15 - 5 .02 ( m . product was obtained by filtration ( 60 . 5 mg, 98 % ) . ‘ H NMR 2H ), 4 . 64 ( s , 2H ), 3 .52 ( d , v = 7 . 0 Hz, 2H ), 1 .48 ( s , 9H ) . 13C (300 MHz, CD , OD ) 8 8 . 32 - 8 . 12 ( m , 2H ) , 8 . 09 - 7 . 92 ( m , 2H ) , NMR ( 126 MHZ. CDC1. ) 8 176 . 7 . 168 . 1 . 160 . 2 . 155 . 7 . 7 .42 - 7 .20 ( m , 4H ) , 4 .62 ( s , 2H ) , 3 .65 - 3 . 41 ( m , 2H ) , 3 . 11 155 . 2 . 132 .9 . 131 . 2 . 131 . 0 . 129 . 5 . 120 . 7 . 120 .4 . 118 . 6 . 25 3 .03 ( m , 1H ) , 2 .53 ( dd , J = 6 . 5 , 1 . 5 Hz, 1H ) , 2 . 16 ( dd , J = 5 . 2 , 118 . 0 , 80 . 9 , 38 . 5 , 37 .4 , 28 . 6 . HRMS (ESI + , m /z ) : calcd for 1 . 5 Hz , 1H ) . 13C NMR (75 MHz, CD , OD ) 8 174 . 7 , 169 .2 , C23H26N304S [ M + H ] * , 440 . 1639; found , 440 . 1634 . 163 . 0 , 159 . 8 , 134 . 8 , 132 . 4 , 130 . 8 , 123 . 8 , 121. 4 , 120 . 0 , 63 . 1 , 36 . 2 , 26 . 9 , 24 . 1 . HRMS (ESI + , m / z ) : Calcd for t- Butyl ( ( 3 - ( 4 - ( 4 - ( (oxiran - 2 - ylmethyl) sulfonyl ) phe C18H , gN30492[ M + H ] * , 404. 0733 ; found , 404 .0746 . noxy ) phenyl) - 1 , 2 , 4 -oxadiazol - 5 - yl) methyl ) carbam 3030 ate (13 ) t- Butyl 4 - (4 -( allylthio )phenoxy )benzylcarbamate ( 15 ) m - CPBA (2 .03 g , 11. 8 mmol) was added in batches to a solution of 12 ( 1 . 04 g , 2 . 36 mmol) in CH ,C1 , ( 8 mL ) A solution of compound 10 ( 4 .98 g , 18 .63 mmol) in THF immersed in an ice -water bath . After completion of the 35 (78 mL) was added dropwise to LiAlH , ( 2 . 12 g , 55 . 89 addition , the ice -water bath was removed and the solution mmol) in THF ( 78 mL ) at 0° C . over a period of 30 min . The was stirred at room temperature for 3 days. Another batch of ice -bath was removed and the reaction mixture was stirred m -CPBA ( 1 .02 g , 5 .89 mmol) was added , and the mixture at room temperature for 1 . 5 h at which point the TLC was stirred at room temperature for an additional 5 d . The showed the reaction to be complete . The solution was cooled suspension was filtered , and the filtrate was diluted with 40 again to ice -water temperature and quenched carefully with CH , C1, and washed with 10 % aqueous sodium thiosulfate , the dropwise addition of 2 . 4 mL water, 2 . 4 mL 15 % aqueous followed by saturated NaHCO , and brine . The organic layer NaOH , and 7 . 2 mL water. The solution was gradually was dried over anhydrous Na SO4, the suspension was warmed to room temperature and stirred for 30 min , filtered filtered , and the solution was concentrated in vacuo . The through a celite pad , extracted with diethyl ether and EtoAc. product was purified by silica gel chromatography (hexanes / 45 The combined organic layer was washed with water and EtOAc , 2: 1 to 1 :1 ) to yield 13 (0 .85 g , 74 % ) . ' H NMR (500 brine , and the solution was dried over anhydrous Na2SO4. MHz, CDC1z) 8 8 .05 ( d , J = 8 .8 Hz, 2H ) , 7 . 87 ( d , J = 8 . 9 Hz, The solvent was evaporated under reduced pressure to give 2H ) , 7 .11 ( d , J = 8 . 9 Hz , 4H ) , 5 . 58 (s , b , 1H ) , 4 .59 (d , J = 5 .8 the crude primary amine, which was used directly in the next Hz , 2H ) , 3 . 38 - 3 . 24 ( m , 3H ) , 2 .79 - 2 . 76 ( m , 1H ) , 2 . 55 - 2 . 34 step . ( m , 1H ) , 1 .42 ( s , 9H ) . 13C NMR ( 126 MHz , CDC12) 8 176 . 9 , 50 To a mixture of amine ( 4 . 3 g , 15 .84 mmol) and ( Boc ) , 173 .9 , 167 .9 , 162 . 1 , 157 .8 , 133 .7 , 131. 0 , 129. 9 , 123 . 5 , (5 .2 g , 23 .77 mmol) in MeOH / CH _ C12 ( 3 : 2 , 150 mL ) , was 120 . 6 , 118 . 7 , 81 . 1, 59 . 9 , 46 . 13 , 46 . 12 , 37 . 5 , 28 . 6 . HRMS added a catalytic amount of iodine (402 mg, 1 .58 mmol, 10 (ESI + , m / z ): calcd for C23H26N30 , S [ M + H ] * , 488 . 1486 ; mol % ) . After stirring the reaction mixture for 24 h at room found , 488 . 1498 . temperature, the solvent was evaporated in vacuo , and ETOAc was added . The solution was washed with 5 % t- Butyl ( ( 3 - ( 4 -( 4 -( ( thiiran - 2 - ylmethyl) sulfonyl ) phe aqueous Na S202 and saturated NaHCO3, and dried over noxy ) phenyl) - 1 , 2 , 4 -oxadiazol - 5 - yl) methyl ) carbam anhydrous Na SO4. The solvent was evaporated in vacuo , ate ( 14 ) and the residue was purified by silica gel chromatography (hexanes / EtoAc, 95 : 5 ) to afford compound 15 ( 3 .21 g , 55 % Thiourea (55 . 3 mg, 0 .73 mmol) was added to a solution 60 in two steps ). ' H NMR ( 300 MHz , CDC12 ) 8 7 . 38 - 7 .32 ( m . of compound 13 ( 161. 1 mg, 0 .33 mmol) in MeOH /CH , C1, 2H ), 7 .31 - 7 .25 (m , 2H ), 6 .99 - 6 .90 ( m , 4H ), 6 . 17 -6 .09 ( m , ( 1 : 1 , 3 mL ), and the resulting mixture was stirred at room 1H ) , 5 .08 -5 .03 (m , 2H ), 4 .85 ( s , b , 1H ) , 4 .29 ( d , J = 3 . 0 Hz, temperature for 24 h . The solvent was removed under 2H ) , 3 .49 -3 .47 ( m , 2H ), 1. 46 (s , 9H ) . 13C NMR (75 MHz, reduced pressure and the residue was partitioned between CDC13 ) & 156 .8 , 156 . 4 , 156 . 1 , 134 .0 , 133 . 2 , 131. 4 , 131. 2 , CH , C1, and water. The organic layer was washed with water 65 129 . 2 , 127 . 2 , 119 . 3 , 117 . 7 , 79 . 7 , 44 . 4 , 38 . 8 , 28 . 6 . HRMS and brine, dried over anhydrous Na2SO4, and filtered . (ESI + , m / z ): calcd for C21H25NNaOzS [ M + Na ] *, 394 . 1447 ; Evaporation of the solvent gave the crude product, which found , 394 . 1472 . US 9 ,951 ,035 B2 55 56 t -Butyl 4 - ( 4 - (( oxiran - 2 -ylmethyl ) sulfonyl ) phenoxy ) 1 - ( 4 - ( 4 - ( ( Thiiran - 2 -ylmethyl ) sulfonyl) phenoxy )phe benzylcarbamate ( 16 ) nyl) - 2 , 3 - bis ( t - butoxycarbonyl) guanidine ( 18 ) Compound 16 was prepared following the same proce - DIEA ( 31 uL , 0 . 17 mmol) was added to a solution of dure as described for the synthesis of compound 13 . Yield , 5 compound 2 (56 . 8 mg, 0 . 16 mmol) and ( Z ) - t - butyl ( ( ( t 36 % . ? H NMR (400 MHz , CDC1, ) 8 7 .91 - 7 .65 ( m , 2H ) , butoxycarbonyl) imino ) ( 1H - pyrazol- 1 - yl) methyl ) carbamate 7 .27 - 7 .25 ( m , 2H ) , 7 . 06 -6 .95 ( m , 4H ), 5 .22 ( s , b , 1H ), 4 .24 ( 44 . 3 mg, 0 . 14 mmol ) in THF ( 0 . 5 mL) . The resulting ( d , J = 5 . 1 Hz, 2H ) , 3 . 44 - 3 .02 ( m , 3H ) , 2 .72 ( dd , J = 8 . 0 , 2 .0 reaction mixture was stirred at room temperature for 24 h . Hz, 1H ), 2 . 39 (dd , J = 4 . 8 , 2 .0 Hz, 1H ), 1 .39 ( s , 9H ). 13C The organic solvent was removed under reduced pressure NMR ( 100 MHZ . CDC1, ) 8 163 . 0 . 154 . 1 . 136 . 6 . 130 .7 . 10 and the residue was purified by preparative TLC ( hexanes! EtOAc, 3 : 1 ) to give 18 (54 . 8 mg, 68 % ) . ' H NMR ( 300 MHz, 129 .5 , 123 . 2 , 120 .7 , 118 . 0 , 117 .8 , 79 .7 , 59 .8 , 46 .04 , 45 .99 , CDC1Z ) 8 11 .64 ( s , 1H ), 10 . 40 ( s , 1H ) , 7 .85 ( d , J = 8 . 7 Hz, 44 .1 , 28 .6 . HRMS (ESI + , m / z ): calcd for C2 H25NNOGS 2H ) , 7 .67 ( d , J = 8 . 7 Hz, 2H ) , 7 .07 ( d , d , J = 8 . 8 Hz, 4H ) , 3 .53 [ M + Na] * , 442. 1295 ; found , 442 . 1292 . ( dd , J = 13 . 9 , 5 . 3 Hz, 1H ) , 3 . 16 (dd , J = 13 . 9 , 8 . 1 Hz, 1H ) , 3 . 09 - 3 .01 ( m , 1H ) , 2 .54 (dd , J = 6 . 1 , 1 . 6 Hz, 1H ) , 2 . 16 (dd , t -Butyl 4 - ( 4 - (( thiiran - 2 - ylmethyl) sulfonyl) phenoxy ) J = 4 .8 , 1. 6 Hz , 1H ), 1. 52 (s , s, 18H ); 13C NMR (75 MHz, benzylcarbamate ( 17 ) CDC12 ) 8 163 . 6 , 163 . 3 , 153 . 8 , 153 . 6 , 151. 5 , 134 . 3 , 132 . 0 , Compound 17 was prepared following the same proce 131 . 0 , 124 . 3 , 121 . 2 , 117 . 8 , 84 . 2 , 80 . 2 , 62 . 9 , 28 . 4 , 28. 4 , 26 . 4 , dure as described for the synthesis of compound 14 . Yield , 20 24564 .6 ;1833 HRMS . found (ESI + 564, m /1836z ): calcd . for C26H34N20 , 52 [ M + H ] ", 58 % . ' H NMR ( 300 MHz , CDC13 ) 87 .85 ( d , J= 8 .7 Hz , 2H ), 7 .33 ( d , J= 8 .7 Hz , 2H ) , 7 .13 -6 .97 ( m , 4H ), 4 .95 ( s, b , 1H ), 1 - (4 -( 4 - (( Thiiran - 2 -ylmethyl ) sulfonyl) phenoxy )phe 4 . 33 ( d , J = 5 . 9 Hz, 2H ), 3 .51 ( dd , J = 14 . 1 , 5 . 7 Hz, 1H ), 3 . 17 nyl) guanidine HCl salt ( 7 ) ( dd , J= 14 . 1 , 7 . 8 Hz, 1H ) , 3 . 11 - 2 . 98 ( m , 1H ) , 2 . 53 ( dd , J = 6 . 3 , 1 .6 Hz , 1H ) , 2 . 15 (dd , J= 5 . 1 , 1 .6 Hz, 1H ) , 1 .46 ( s , 9H ). 13C 25 Hydrogen chloride gas was passed through a solution of NMR (75 MHz, CDC1z ) & 163. 1 , 156 . 0 , 154 . 0 , 136 . 3 , 132. 1 , 18 ( 37 . 1 mg) in CH2C1_ /EtOAC ( 1 : 1 , 3 mL ) under ice -water 130 . 8 , 129 . 4 , 120 . 7 , 117 . 8 , 79 . 8 , 62 . 8 , 44. 1 , 28 . 5 , 26 . 2 , 24 . 4 . temperature for 3 min . The organic solvent was then HRMS (ESI + , m / z ) : calcd for C2 H25NNaO , S2[ M + Na ] ", removed under reduced pressure to give 7 ( 37 . 1 mg, 95 % ) . 458 . 1066 ; found , 458 . 1089 . H NMR (500 MHz, DMSO -d ) 89. 92 (s , 1H ), 7. 91 (d , J = 8 . 8 30 Hz, 2H ) , 7 .50 ( s , 3H ) , 7 . 34 ( d , J = 8 . 8 Hz, 2H ) , 7 . 28 - 7 . 16 ( m , (4 - ( 4 - ( ( Thiiran - 2 - ylmethyl) sulfonyl) phenoxy ) phe 4H ) , 3 . 66 ( m , 2H ) , 2 . 99 ( m , 1H ) , 2 .56 (dd , = 6 . 0 , 1 . 0 Hz, 1H ) , nyl) methanamine HC1 salt ( 5 ) 2 . 16 (dd , J= 5 . 0 , 1 .0 Hz, 1H ). 13C NMR ( 126 MHz, DMSO d ) 8 162 .5 , 157 . 0 , 153 .7 , 133. 3 , 132. 7 , 131. 5, 128 .1 , 122 . 2 , Compound 5 was prepared following the same procedureure 118 . 4 , 61 .3 , 27 . 7 , 24 .6 . HRMS ( ESI + ): calcd for as described for the synthesis of compound 4 Yield 98 % 35 C16H18N303S2 [ M + H ] * , 364 .0784 ; found , 364 .0770 . Enzyme Inhibition Studies . ' H NMR ( 300 MHz , CD2OD ) 8 7 . 93 ( d , J = 8 .6 Hz, 2H ) , 7 .58 Human recombinant active MMP - 2 and MMP - 7 , and the ( d , J = 8 .6 Hz, 2H ), 7 . 19 (d , d , J = 8 . 4 Hz, 4H ), 4 . 16 ( s, 2H ), catalytic domains ofMMP -3 and MMP - 14 /MT1 -MMP were 3 .57 -3 .43 ( m , 2H ), 3. 11 - 2 .99 ( m , 1H ), 2 .52 (dd , J = 6 .3 , 1 .4 purchased from EMD Chemicals , Inc. ( San Diego , Calif ., Hz, 1H ), 2 .14 (dd , J = 5 . 1, 1. 4 Hz , 1H ). 13C NMR (75 MHz·, 40 USA ); human recombinant catalytic domains of MMP - 1 , CD2OD ) 8 162 .5 , 156 .2 , 133 .0 , 131 .3 , 131. 1, 130 .0 , 120 . 7 , MMP - 8 , and MMP - 9 were purchased from Enzo Life Sci 118 . 2 , 62 . 0 , 42 . 6 , 25 . 8 , 23 . 0 . HRMS ( ESI + , m / z ) : calcd for ences , Inc . (Farmingdale , N . Y . , USA ) ; human recombinant C16H , NO2S2 [ M + H ] * , 336 .0723 ; found , 336 .0709 . active ADAM9 and ADAM10 were purchased from R & D Systems (Minneapolis , Minn . , USA ) . Fluorogenic substrates N - ( 4 - ( 4 -( ( Thiiran - 2 - ylmethyl) sulfonyl) phenoxy ) 45 MOCAC- Pro - Leu -Gly - Leu -A2pr ( Dnp )- Ala- Arg - NH2 ( for benzyl) acetamide (6 ) MMP- 2, MMP - 7, MMP- 9 and MMP -14 ) and MOCAC- Arg Pro - Lys - Pro - Val -Glu -Nva - Trp - Arg - Lys (Dnp )- NH , ( for Acetyl chloride (43 . 6 uL , 0 .61 mmol) in THF ( 0 . 5 mL ) MMP - 3 ) were purchased from Peptides International (Lou was slowly added to a cooled solution of 5 (69 mg, 0 . 19 isville , Ky. , USA ) ; Mca -KPLGL - Dpa - AR -NH , ( for MMP mmol ) , triethylamine (85 . 4 UL , 0 .61 mmol) in THF ( 1 . 0 50 1 , MMP - 8 and ADAM10 ) and Mca- PLAQAV - Dpa -RSSSR mL ) . The reaction mixture was stirred for 3 h under ice NH , ( for ADAM9) were purchased from R & D Systems water bath , warmed to room temperature , and allowed to stir (Minneapolis , Minn . , USA ). The K , values for MMP - 2 , overnight. The reaction was quenched with saturated MMP - 9 and MMP - 14 were as previously reported by Gooyit NaHCO3 and the product was extracted with CH ,C12 . The et al ., J . Med. Chem . 2013 , 56 , 8139 -8150 . Inhibitor stock combined organic layers were dried over anhydrous 55 solutions ( 10 mm ) were prepared fresh in DMSO before Na2SO4, filtered , and evaporated to dryness . The residue enzyme inhibition assays . We followed the same methodol was purified by preparative TLC ( hexanes / EtoAc/MeOH , ogy for enzyme inhibition studies as reported by Page 14 :85 : 1) to give 6 (45 .8 mg, 65 % ). ' H NMR (500 MHz , McCaw et al. (Nat . Rev. Mol. Cell Biol. 2007 , 8 , 221 -233 ) . CDC1z ) 8 7 .87 ( d , J = 8 . 5 Hz, 2H ) , 7 . 36 ( d , J = 8 . 5 Hz, 2H ) , Enzyme inhibition studies were carried out using a Cary 7 .08 ( d , d , J = 8 . 6 Hz, 4H ) , 6 .03 ( s , b , 1H ) , 4 .47 ( d , J = 5 . 9 Hz, 60 Eclipse fluorescence spectrophotometer ( Varian , Walnut 2H ) , 3 .52 ( dd , = 14 . 2 , 5 . 7 Hz, 1H ) , 3 . 21 (dd , - 14 . 2 , 7 . 7 Hz, Creek , Calif . , USA ) . Compounds 4 , 5 , 6 and 7 were stable 1H ), 3 . 14 - 2 . 98 ( m , 1H ) , 2 . 56 ( dd , J = 6 . 2 , 1 . 6 Hz, 1H ) , 2 . 18 in the buffers that were used in the kinetic assays. ( dd , = 5 . 1 , 1 .6 Hz, 1H ) , 2 .07 ( s , 3H ) . 13C NMR ( 126 MHz, Computational Analysis . CDC13 ) 8 170 . 1 , 163. 0 , 154. 5 , 135 . 8 , 132. 6 , 130 . 9 , 129. 9 , MMP - 2 protein coordinates were obtained from our pre 120 . 8 , 118 . 0 , 63 . 0 , 43. 3 , 26 . 2 , 24 . 4 , 23 . 4 . HRMS ( ESI + , 65 vious QMMM study ( Zhou et al. , J . Chem . Theory Comput . m / z ) : calcd for C18H2. NO S2 [ M + H ] * , 378 . 0828 ; found , 2010 , 6 , 3580 -3587 ) . Coordinates ofMMP - 9 and MMP - 14 378 .0836 . were downloaded from the Protein Data Bank ( PDB codes US 9 , 951, 035 B2 57 58 of 1GKC and 3MA2 , respectively ) and prepared using 319 - 182 for 5 , 378 – 182 for 6 , 364 - > 227 for 7, and Protein Preparation Wizard via Maestro v 9 . 3 . 5 ( Schrodinger 300 - 93 for internal standard 19 . LLC , Portland , Oreg . , USA ). Compounds were prepared using LigPrep v2 .55 . Molecular docking of the compounds to the catalytic site of MMPs was carried out with Glide 5 5 v5 . 8 , implementing 1- A core restraints of the thiirane group and scored with Standard Precision ( Friesner et al. , J . Med . Chem . 2004 , 47 , 1739 - 1749 ) . Animals . N Mice (male CD - 1 , 6 - 7 weeks old , - 30 g body weight, specific pathogen free ) were purchased from Charles River Quantification of the compounds in plasma and brain was Laboratories, Inc . (Wilmington , Mass ., USA ) . Mice were obtained using peak - area ratios of the compounds to the fed Teklad 2019 Extruded Rodent Diet (Harlan , Madison , internal standard , and the linear- regression parameters Wis . , USA ) and provided with water ad libitum . Animals 15 obtained from the calibration curves. The coefficients of were housed in polycarbonate shoebox cages containing determination ( R ) were > 0 .99 , and the assays were linear Bed - o ' Cobs 14 " ( The Andersons Inc .. Maumee . Ohio / Al- up to concentrations of 100 uM . Pharmacokinetic Parameters . pha -dri (Sheperd Specialty Papers , Inc ., Richland , Mich .) The methodology for the calculation of pharmacokinetic bedding under a 12 h light/ 12 h dark cycle at 72 + 2° F . parameters listed in Table 3 is as reported by Gooyit et al. (J . Animal Dosing and Sample Collection . 20 Med . Chem . 2013 , 56 , 8139 - 8150 ) . Compounds 4 , 5 , 6 , and 7 were formulated as a solution at a concentration of 1 .25 mg/mL . Compounds 4 , 5 , and 7 Example 2 . Selective Water- Soluble and were dissolved in 73 % water /20 % propylene glycol/ 7 % Slow -Binding Matrix Metalloproteinase Inhibitors DMSO . Compound 6 was dissolved in 61 % water / 27 % for Traumatic Brain Injury Therapy, for propylene glycol/ 12 % DMSO . Mice were given a single 25 Neurological Diseases , and for Wound Healing 120 - uL iv dose of compounds 4 , 5 , 6 , or 7 ( equivalent to 5 Therapy mg/ kg , n = 3 per time point) . The sterilization of the dosing solutions and the procedure for collection of both plasma Traumatic brain injury ( TBI) is a devastating disease and brain as reported by Gooyit et al. ( J. Med . Chem . 2013 , caused by primary and secondary injuries that can lead to a 56 , 8139 - 8150 ) . 30 chronic disease process characterized by persistent cognitive Sample Analysis . deficits . This disease process starts with biochemical Procedures for the preparation of plasma and brain changes that result in brain cell damage and cell death over samples , as well as calibration curves for quantification months to years later . Matrix metalloproteinase - 9 (MMP - 9 ) analysis , were as described by Gooyit et al. ( J . Med . Chem . is at the top of the biochemical cascade of events that lead 2013 , 56 , 8139 - 8150 ) . Samples were analyzed by ultraper - 35 to secondary injury . Wehave shown that selective inhibition formance liquid chromatography (UPLC )/ ( + ) electrospray of MMP - 9 attenuates secondary damage resulting from TBI. ionization (ESI ) - multiple - reaction monitoring (MRM ) with We have created water- soluble selective MMP - 9 inhibitors a reversed phase C18 column (Acclaim® RSLC 120 C18 , to address the therapeutic need in the area of TBI treatment 2 . 2 um , 120 Å , 2 . 1x100 mm , Dionex , Sunnyvale , Calif ., ( Table 2 . 1 ) . The compounds ND - 336 and ND - 378 cross the USA ) . The chromatographic and mass spectrometric condi - 40 blood - brain barrier and achieve therapeutic concentrations tions were as previously reported by Gooyit et al . ( J. Med. in the brain (FIG . 5 ). TABLE 2 . 1 H?N QmwW ND - 336 ( 5 ) ND - 378 (6 ) Ki (MMP - 2 ) 0 .085 + 0 . 001 uM ( slow - binding ) Ki (MMP - 2 ) 0 . 234 + 0 . 006 UM ( slow -binding ) Ki (MMP - 9 ) 0 . 150 + 0 . 005 uM ( slow - binding ) Ki (MMP - 9 ) 23 % inhibition at 50 UM Ki (MMP - 14 ) 0 . 120 + 0 .005 UM (slow -binding ) Ki (MMP - 14 ) 23 % inhibition at 50 UM Ki (MMP - 8 ) 7 . 72 = 0 .07 UM ( linear noncompetitive ) Ki (MMP - 8 ) 0 .692 = 0 .042 uM (linear competitive ) Ki (MMP - 1 ) 4 % inhibition at 100 UM Ki (MMP - 1 ) 37 % inhibition at 100 UM Ki (MMP - 3 ) 23 % inhibition at 100 UM Ki (MMP - 3 ) 14 % inhibition at 100 UM Ki (MMP - 7 ) 1 % inhibition at 100 uM Ki (MMP - 7 ) 4 . 7 % inhibition at 100 UM Ki (ADAM - 9 ) 31 % inhibition at 100 UM Ki (ADAM - 9 ) 31 % inhibition at 100 uM Ki ( ADAM - 10 ) 14 % inhibition at 100 UM Ki ( ADAM - 10 ) 8 % inhibition at 100 UM Water Solubility 4 . 9 mg/ mL Water Solubility 0 .025 mg/mL

Chem . 2013 , 56 , 8139 -8150 ) , except for the following : the Gelatinases (matrix metalloproteinases (MMP ) - 2 and 9 ) capillary voltage, cone voltage , extractor voltage , and RF play important roles in the pathology ofmany neurological lens voltage were set at 4 . 6 kV, 25 V , 3 V , and 0 . 1 V , 65 diseases, as well as in wound healing . A major challenge to respectively ; the cone gas - flow rate was set at 50 L /h the development of therapeutics for the treatment of neuro (nitrogen ) . The MRM transitions were 404 - > 209 for 4 , logical diseases is the inability of > 98 % of small -molecule US 9 ,951 , 035 B2 59 60 drugs to cross the blood -brain barrier ( BBB ) and achieve SB -3CT , it crosses the BBB , and achieves therapeutic therapeutic concentrations in the brain . concentrations in the brain . Furthermore , we have SB - 3CT ( compound 1 ) is a selective slow - binding and shown that MMP- 9 cleaves tau and that ND - 336 prevents potent inhibitor of gelatinases that shows efficacy in animal cleavage of tau (FIG . 6 ) . Because cleaved tau can lead models of neurological diseases . However , this compound is 5 to chronic neurodegeneration after traumatic brain injury poorly water - soluble . We synthesized and evaluated p - amin - ( TBI) and plays an important role in the pathology of omethyloxadiazol ( 4 ) , p - aminomethyl (5 , ND - 336 ) , p - acet Alzheimer' s disease and chronic traumatic encephalopathy, amidomethyl ( 6 , ND - 378 ) , and p - guanidino ( 7 ) compounds ND - 336 can be used to reverse cognitive dysfunction fol as improvements on compound 1 . The compounds are 10 - to lowing TBI. TABLE 2 .2 MMP -9 Ki MMP- 8 Ki (nM ) (nM ) 870 + 110 2600 - 400

ND -322 (2 ) 150 + 10 7700 + 100

H?N .

ND -336 (5 ) O 180 + 30 13000 + 2000

Me01

ND - 380

14 ,000 - fold more water - soluble than 1, retained slow -bind - ND -378 is also a selective MMP- 2 inhibitor that also ing inhibition behavior towards MMP- 2 , and cross the BBB . 40* crosses the BBB . Recent efforts in defining the role of The p -acetamidomethyl analog ( compound 6 ) is a selec - MMPs in cancer have correlated increased expression of tive nanomolar slow -binding inhibitor of MMP - 2 , which MMP - 2 with aggressive breast cancer, malignant prostate does not inhibit the closely related MMP -9 or MMP - 14 . cancer , pancreatic cancer, gastric cancer, brain metastasis Because of the slow dissociation of compound 6 from the 45 and melanoma, and lung cancer brain metastasis . Tumors target MMP - 2 ( residence time of 6 bound to MMP - 2 is expressing MMP - 2 have increased vasculature at the brain 18 . 2 + 0 . 4 min ), it results in sustained inhibition of MMP - 2 tumor interface, indicating that MMP - 2 plays a role in even when concentrations of 6 fall below the Ki value . This enhancing invasion and vascularization within the central inhibitor is a useful tool in therapeutic intervention and in nervous system . Therefore , selective MMP - 2 inhibition can investigations of the role of MMP - 2 in neurological dis - 50 be a targeted therapy for the treatment of brain metastasis . eases. The p -aminomethyl derivative ( compound 5 ) is a water- soluble nanomolar slow - binding inhibitor of MMP - 2 , Example 3 . MMP - 9 is Detrimental, MMP- 8 is MMP- 9 , and MMP - 14 and has residence times for inhibition Beneficial, and MMP - 2 is not Involved in Diabetic of these enzymes 6 - to 7 - fold longer than those of the tissue Wound Healing inhibitors of metalloproteinase 1 or 2 ( TIMP - 1 or TIMP - 2 ) 55 bound to MMP- 9 , protein inhibitors that have evolved for The incidence of diabetes mellitus is increasing . In 2010 , the purpose of regulating MMPs . 8 . 3 % of the population in the United States had diabetes Chronic wounds are a complication of diabetes. We (approximately 25 . 8 million Americans) . Two years later , previously showed that MMP- 9 is detrimental to wound the percentage increased to 9 . 3 % (29 . 1 million people ) . A healing , while MMP - 8 is involved in repair of wounds. The 60 complication of diabetes is the inability of wounds to heal, p -aminomethyl compound ND - 336 has better selectivity which results in greater than 60 % of non - traumatic limb towards MMP - 9 than the p -amino compound ND -322 amputations occurring in individuals with diabetes. The ( Table 2 . 2 ) . In experiments involving a mouse model of number of lower - limb amputations was 65 ,700 in 2006 , diabetic wound healing , ND - 336 showed significantly better increasing to 73 , 000 in 2010 . efficacy than ND - 322 ( FIGS. 3 and 4 ) . 65 Wound healing involves four stages: hemostasis, inflam ND -336 is > 2 ,000 - fold more water - soluble than SB -3CT , mation , proliferation , and remodeling . During hemostasis , 4 - fold more potent towards inhibition of MMP- 9 than blood vessels are constricted and a fibrin clot is formed . The US 9 ,951 ,035 B2 61 62 fibrin clot releases pro - inflammatory cytokines and growth vs 75 + 9 % , n = 6 , p = 0 .5 , FIGS . 7a and 76 ). Partial re - epithe factors , followed by infiltration of neutrophils , macro - lialization was observed in both MMP - 2 - inhibitor -treated phages, and lymphocytes. Neutrophils release reactive oxy and vehicle -treated groups (FIG . 7c ) . Hence, selective inhi gen species and matrix metalloproteinases (MMPs ), while bition of MMP- 2 has no effect in diabetic wound healing . macrophages induce apoptotic cells . As apoptotic cells are 5 This study confirmed that MMP - 2 plays no role in diabetic cleared , angiogenesis is stimulated to promote re - epitheli wound healing . Unfortunately , the role of MMP - 2 in dia alization . Collagen is produced , as well as extracellular betic wound healing could not be confirmed with a study in matrix (ECM ) components , leading to remodeling and MMP - 2 knockout mice , as ablation of MMP - 2 results in a wound closure. Any disruption , abnormality , or prolongation compensatory increase in MMP - 9 , making it difficult to 10 ascertain its role by this method . in this process leads to delayed wound healing or a chronic We had previously shown thatMMP - 9 was detrimental to wound . diabetic wound healing by selective chemical inhibition MMPs are a family of 26 proteinases that are zinc (Gooyit et al. , J . Med . Chem . (2013 ) 56 , 8139 -8150 ) . To dependent and degrade the ECM . MMPs play an important confirm the detrimental role of MMP - 9 in diabetic wound role in wound healing , cleaving excess ECM , as well as 15 healing , we used MMP - 9 knockout mice . We induced dia proteolytically process chemokines and cytokines . Nor betes in the animals with streptozotocin . Streptozotocin is mally , MMP activity is regulated by complexation with highly toxic to the insulin -producing beta cells of the tissue inhibitors of matrix metalloproteinases ( TIMPs) . pancreas and produces an animal model for type 1 diabetes However , in chronic wounds MMP activity is dysregulated (Szkudelski , Physiol. Res. (2001 ) 50 , 537 - 546 ) . Streptozo and the elevated levels of MMPs contribute to excessive 20 tocin treatment ( 150 mg/ kg given intraperitoneally ) reliably degradation of the ECM and lead to a stalled wound healing induces diabetes in two weeks , as confirmed with fasting process. blood glucose levels of > 300 mg/ dL . Diabetes was also We recently reported on the identification of MMP - 8 and induced with streptozotocin in wild -type mice of the same MMP- 9 in wounds of diabetic mice (Gooyit et al ., ACS background as the controls . Significant differences in wound Chem . Biol. 2014 , 9 , 505 - 510 ) . We used an inhibitor 25 healing between MMP - 9 knockout and wild - type streptozo tethered resin that binds only to active MMPs, to the tocin - induced diabetic mice were observed on day 7 exclusion of MMP zymogens and TIMP - complexed MMPs. ( 76 + 14 % vs 58 + 24 % , n = 14 , p < 0 . 05 , FIGS . 8a and 8b ) . On Using this resin we did not detect MMP - 2 , however we day 14 , wound closure in MMP- 9 knockouts was lower than observed an active MMP - 2 band by gelatin zymography, in wild - type mice , although it was not statistically signifi suggesting that MMP- 2 was in complex with TIMP. MMP- 2 30 cant (95 6 % vs 89 + 9 % , n = 7 , p = 0 .15 , FIG . 8a ). Partial and MMP- 9 have been reported in wounds of diabetic re -epithelialization was seen in wild - type streptozotocin patients and diabetic mice . In these studies, the levels of the induced diabetic mice (FIG . 86 , upper right) , however gelatinases were determined by ELISA , gelatin zymogra re - epithelialization was complete in MMP - 9 knockout strep phy, mRNA , and Western blot. However , these methods tozotocin - induced diabetic mice ( FIG . 8b , lower right) . cannot distinguish between active gelatinases and TIMP 35, In -situ gelatin zymography showed the absence of gelatinase complexed gelatinases (inactive forms) . Sodium dodecyl activity in wounds ofMMP - 9 knockout mice , as evidenced sulfate is used in gelatin zymography , which denatures the by considerable decrease in fluorescence intensity (FIG . 8c , inactive TIMP -MMP complex and results in the appearance lower left ) . Altogether, we conclude that gene ablation of of active MMP bands . As such , the MMP - 2 reported in MMP - 9 accelerates wound healing in diabetic mice . These wounds of diabetic patients and diabetic mice coulda inin lactfact 40an resultsres confirmed that MMP -9 is detrimental to wound be inactive MMP - 2 complexed with TIMP. healing . We recently designed and synthesized a selective MMP - 2 We used a uniquely versatile inhibitor- tethered resin for inhibitor ( compound 1x ) ; this compound inhibits MMP - 2 as identification of active MMP - 8 and MMP - 9 in both diabetic a slow -binding inhibitor with a K , value of 440 + 60 nM and and non - diabetic wounds, and showed that the levels of the poorly inhibits or does not inhibit other MMPs, including8 45 latter were elevated at statistically significant levels only in MMP -8 (K ;= 17 ,000 + 2 ,000 nM ) and MMP - 9 (28 % inhibi diabetic wounds. Working with the hypothesis that MMP - 9 tion at 50 ,000 nM ) (Gooyit et al. , J . Med . Chem . ( 2013 ) 56 , is detrimental to healing of diabetic wounds , but that 8139 - 8150 ) . The residence time for compound 1x bound to MMP - 8 likely plays a beneficial effect, we inhibited MMP - 9 MMP- 2 is 24 . 5 min , which is significantly longer than those selectively by the use of ND -322 . The diabetic wounds of TIMP- 1 and TIMP - 2 bound to MMP- 2 of 6 . 9 and 10 . 4 50 healed more rapidly in a process that involved re -epitheli min , respectively . Thus, compound 1x is more effective at alization of the wounds, as is the case for the non - diabetic inhibiting MMP- 2 than TIMPs . wounds in wild - type mice . In addition , apoptosis was sig nificantly attenuated . We also used a selective MMP- 8 1x inhibitor and showed that healing of the diabetic wounds 5 was delayed , accompanied by decreased re - epithelialization , and undiminished apoptosis . The present study reveals a beneficial effect of selective inhibition of MMP- 9 in healing of diabetic wounds. Whereas the use of the selective inhibi tor ND - 322 does not show any effect on non -diabetic IZ s 60 wounds neither detrimental nor beneficial — it is intriguing that the use of the broad - spectrum MMP inhibitor ilomastat ( also known as GM - 6001) in non - diabetic wounds in rats , We used compound 1x to determine the effect of inhib - pigs , and humans delayed wound closure and diminished iting MMP - 2 on diabetic wound healing . No significant epithelialization . These findings reveal that broad inhibition differences were observed in MMP - 2 inhibitor- treated and 65 of the “ good ” and the “ bad ” MMPs simultaneously is vehicle groups ( day 7 : 23 - 16 % vs 18 21 % , n = 12 , p = 0 . 7 ; detrimental to the wound -healing process . Clinical manage day 10 : 45 + 11 % vs 50 + 11 % , n = 6 , p = 0 . 5 ; day 14 : 70 15 % ment of diabetic wounds presently involves merely debride US 9 , 951, 035 B2 63 64 ment of the wound and attempts at keeping it clean and free Cloning , Purification , and Kinetic Analysis of Musmus of infection . The selective MMP- 9 inhibition strategy that culus MMP- 8 . we have disclosed here is a first potential pharmacological The catalytic domain ( 304 - 852 bp ) of MMP - 8 was opti intervention in treatment of diabetic wounds, which holds mized for expression in Escherichia coli and synthesized great promise in addressing an unmet medical need . 5 from GenScript (Piscataway , N . J .) with unique Ndel and Methods Xhol restriction sites flanking the gene at the 5 ' and 3 ' Synthesis of Compound 1x . termini, respectively . The gene was then cloned into vector PET28a . The construct was transformed into E . coli DH5a The selective MMP - 2 inhibitor ( compound 1 ) was pre and verified for the correct insert using Ndel and Xhol pared by our methodology as described earlier (Gooyit et al ., 10 double digestion of extracted plasmids and further verified J . Med . Chem . ( 2013 ) 56 , 8139 -8150 ) and dissolved in 20 % by sequencing of both DNA strands . The purification pro DMSO / 80 % propylene glycol at a concentration of 5 . 0 cedures of recombinant MMP - 8 were modified from previ mg/ mL . The dosing solution and respective vehicle solution ously published methods ( Tkalcevic et al. , Toxicol. Pathol. were sterilized by passage through a 0 . 2 um , 13 mm diam (2009 ) 37 , 183 - 192 ) . The verified construct was transformed eter PTFE membrane connected to an Acrodisc syringe filter 15 into E . coli BL21 (DE3 ) cells . The expression of MMP - 8 (Pall Life Sciences ) . was induced by addition of 0 . 5 mM isopropyl B - D - 1 - thio Animals . galactopyranoside at 20° C . Cells were harvested 20 hours Female mice were obtained from the Jackson Laboratory later, re- suspended in buffer A ( 200 mM NaCl, 20 mM and provided with Laboratory 5001 Rodent Diet ( PMI) and HEPES , PH 7 . 5 ) , and sonicated on ice . The lysed cells were water ad libitum . Mice were kept in polycarbonate shoebox 20 centrifuged for 5 min at 20 ,000 g . The pellet was gently cages with hardwood bedding at 72 + 2° F . and a 12 : 12 h washed with buffer A to remove cellular debris and was light/ dark cycle . suspended in buffer A , centrifuged for 30 min at 20 , 000 g , Mouse Diabetic Wound Model. and re- suspended in buffer B ( 6 M urea, 50 mM Tris ( pH The excisional mouse diabetic model validated by Sulli - 8 . 5 ) , 5 uM ZnC1 ,) . Undissolved debris was removed by van et al . was used (Sullivan et al ., Plast . Reconstr. Surg . 25 30 -min centrifugation at 20 ,000 g . The supernatant was ( 2004 ) 113 , 953 - 960 ) . The procedure for inflicting the loaded onto a Q Sepharose column equilibrated in buffer B . wounds ( single 8 mm wound on the dorsal region ) was the linear gradient of 0 to 1 M NaCl was used to elute the same as described previously (Gooyit et al. , ACS Chem . protein . The fractions with the desired protein were pooled Biol . (2014 ) 9 , 505 -510 ). Wounds were photographed and and prepared for dialysis by diluting the protein to an covered with a sterile 3M TegadermTM transparent dressing 30 absorbance of 0 . 3 at 280 nm and increasing the concentra (Butler Schein Animal Health , Inc .) . tion of urea from 6 M to 8 M and the concentration of ZnCl2 MMP - 2 Inhibitor Study . from 5 uM to 50 °M . Female diabetic db /db (BKS . Cg -Dock7m + / + Leprdb /J , Three steps of dialysis were performed . First, dialysis was 8 -weeks old , 38 + 3 g , n = 24 ) were used for this study . performed twice in 1 : 5 protein to buffer C (50 mM Tris ( pH Wounds were inflicted as described . Treatment with the 35 8 . 0 ) , 200 mM NaCl, 100 mM glycine , 10 mM CaCl ,, 3 mM MMP- 2 inhibitor (50 uL of 5 . 0 mg /mL in 80 % propylene NaN3 , 50 UM ZnCl2 ), then in 2 L of buffer D (50 mM Tris glycol/ 20 % DMSO , equivalent to 0 .25 mg per wound ) or ( pH7. 5 ) , 10 mM CaCl2 ) . The protein solution was concen vehicle (50 uL of 80 % propylene glycol/ 20 % DMSO ) was trated with Macrosep® Centrifugal Filters ( Pall Life Sci started one day after wound infliction and continued once a ences ) . The purity of the protein was determined to be > 95 % day for 14 days . Digital photographs of wounds were taken 40 by SDS - PAGE ( FIG . 11 ) . The enzyme concentration was on days 0 , 7 , 10 , and 14 while the animals were under evaluated spectrophotometrically using the extinction coef isoflurane anesthesia . On day 14 , all mice were sacrificed . ficient predicted by ProtParam (Gasteiger et al. , Protein The wounds were excised , embedded in optimal cutting identification and analysis tools on the ExPASy Server , in temperature (OCT ) compound , and cryosectioned for histo The Proteomics Protocols Handbook, 2005 , Walker , J . M . , logical evaluation . 45 Ed ., pp 571 -607 , Humana Press ) ( A£280 = 19681 .6 M MMP- 9 Knockout Study. cm ) . Aliquots of the concentrated protein were stored in 50 Female MMP- 9 knockout mice ( B6 .FVB (Cg ) - mM Tris (pH 7 . 5 ) , 5 mM CaCl , , 300 mM NaCl, 20 uM Mmp9tmlTvu / J , 8 -weeks old , 19 + 2 g , n = 14 ) and wild - type ZnCl2, 0 . 5 % ( w / v ) Brij- 35, 30 % glycerol at - 80° C . mice (C57BLKS /6J , 8 -weeks old , 19 + 2 g , n = 14 , same The K . , kar, and Vw values for the reaction of MMP- 8 background as MMP - 9 knockoutmice ) were used . The mice 50 with the fluorogenic substrate Mca -KPLGL -Dpa - AR -NH2 were acclimated to the study room for one week prior to ( R & D Systems ) were evaluated in reaction buffer ( 50 mM commencement of the study. Diabetes was induced by Tris (pH7 . 5 ) , 10 mM CaCl2 , 150 mM NaCl, 0 .05 % ( w / v ) intraperitoneal injection of streptozotocin (Sigma ) at 150 Brij- 35 ) . The substrate was prepared as a 1 mM stock mg/ kg . Streptozotocin was dissolved in 100 mM sodium solution in DMSO . Substrate hydrolysis was monitored with citrate buffer (pH 4 . 5 ) and administered within 15 min after 55 excitation at 325 nm and emission at 393 nm with a Cary preparation . After streptozotocin treatment, the mice were Eclipse fluorescence spectrophotometer ( Varian ) . A standard housed in disposable cages and given 10 % sucrose water to curve was constructed at various concentrations of the full drink for two days. The fasting blood glucose levels were hydrolyzed substrate for calibration . Initial velocities were determined two days after streptozotocin treatment. Animals obtained from plots of fluorescence versus time. The slope with blood glucose greater than 300 mg/ dL were considered 60 from these plots was divided by the fluorescence change diabetic . Animals with blood glucose less than 300 mg /dL corresponding to complete hydrolysis and then multiplied received a second dose of streptozotocin one week later . by the substrate concentration to obtain initial velocity in Wounds were inflicted as described and digital photographs units of second ( s - ?) . The parameters keat and Km were were taken on days 0 , 7 , and 14 . Mice (n = 7 per group ) were determined by nonlinear fitting of the initial velocities, at sacrificed on days 7 and 14 , the wounds were excised , 65 varying concentrations of the substrate , using the Michaelis embedded in OCT compound , and cryosectioned for histo - Menten equation . Kinetic analysis resulted in K , = 4 . 5 + 0 . 5 logical evaluation . UM , Vmax = 1 .07 + 0 .03 nM s - 1, kcat = 0 .54 + 0 .02 s - 1 , kcal US 9 ,951 ,035 B2 65 66 Km = 1. 19x10 % M - 's - 1, activity = 926 pmol/ min /ug . The cata lytic efficiency of the purified MMP - 8 was high compared to (i ) Tablet 1 mg/ tablet other commercial available MMPs . Compound X ' 100. 0 Exogenous MMP -8 Study . Lactose 77. 5 Female diabetic db / db (BKS . Cg - Dock7m + / + Leprdb / J , 5 Povidone 15 . 0 Croscarmellose sodium 12 . 0 8 - weeks old , 38 + 3 g , n = 24 ) were used for this study . Microcrystalline cellulose 92. 5 Wounds were inflicted and the following day the wounds Magnesium stearate 3 . 0 were treated topically with MMP - 8 (50 uL of 20 ug /mL 129983 MMP - 8 in reaction buffer ) or vehicle ( 50 uL reaction buffer ) 300 .0 once a day for 14 days . The reaction buffer consisted of 50 10 mM Tris (pH 7 . 5 ) , 10 mM CaCl2 , 150 mM NaCl, and 0 .05 % ( ii ) Tablet 2 mg/ tablet ( w / v ) Brij -35 . Digital photographs of the wounds were taken " Compound X ' 20 . 0 on days 0 , 7 , 10 , and 14 while animals were under isoflurane Microcrystalline cellulose 410 . 0 anesthesia . On days 7 and 14 , 12 mice ( n = 6 per group ) were Starch 50 . 0 sacrificed . The wounds were excised , embedded in OCT 15 Sodium starch glycolate 15 . 0 compound , and cryosectioned for histological evaluation . Magnesium stearate 5 . 0 Wound Measurements . 500 . 0 Photographs were taken using the procedure described by Gooyit et al. ( ACS Chem Biol. ( 2014 ) 9 , 505 -510 ) at a fixed ( iii ) Capsule mg/ capsule distance above the wounds and calibrated using a ruler 20 ' Compound X 10 . 0 included in the photographic frame. Image J 1. 48c software Colloidal silicon dioxide 1 . 5 was used to calculate wound areas. The percentage ofwound Lactose 465 . 5 closure was calculated from the wound area on the specified Pregelatinized starch 120 . 0 day relative to that on day 0 . Magnesium stearate 3 . 0 Histological Evaluation , Apoptosis Detection , and In -Situ 25 600 . 0 Zymography. Fresh wound tissue was harvested and embedded in OCT (iv ) Injection 1 (1 mg/ mL ) mg/mL compound . Subsequently , the tissue was cryosectioned for Compound X ’ ( free acid form ) 1 . 0 H & E staining ( 12 - um thickness ) and for apoptosis assays Dibasic sodium phosphate 12 . 0 and in - situ zymography ( 8 -um thickness ) . Re -epithelializa - 30 Monobasic sodium phosphate 0 . 7 Sodium chloride 4 . 5 tion was assessed on a Nikon Eclipse 90i Fluorescent 1 .0N Sodium hydroxide solution q . . Microscope (Nikon Instruments Inc .) as described earlier (pH adjustment to 7 . 0 - 7 . 5 ) ( Tkalcevic et al ., Toxicol. Pathol. (2009 ) 37 , 183 - 192 ). Water for injection q .s . ad 1 mL Apoptosis was evaluated using Derma TACSTM Apoptosis detection kits ( Trevigen , Inc . ) according to the manufactur - 35 ( v ) Injection 2 ( 10 mg/ mL ) mg/mL er ' s instructions . The protocol of in -situ zymography was " Compound X ’ ( free acid form ) 10 . 0 adapted from known methods. Gelatinolytic activity was Monobasic sodium phosphate 0 . 3 detected in unfixed cryostat sections using DQ - gelatin as a Dibasic sodium phosphate 1 . 1 Polyethylene glycol 400 200 . 0 substrate . DQ - gelatin was prepared at a concentration of 20 0 . 1N Sodium hydroxide solution q . s . ug /mL in Tris -buffered saline ( TBS ) buffer ( 50 mM TBS pH 40 (pH adjustment to 7 . 0 - 7 . 5 ) 7 . 6 , 1 mM CaCl2 ) . A general MMP inhibitor in 0 . 5 mM Water for injection q . s . ad 1 mL EDTA was prepared as a negative control. Cryostat sections were air - dried for 10 min and incubated in the substrate ( vi) Aerosol mg/ can mixture in the presence and absence of EDTA for 1 hr at ' Compound X 20 room temperature . After the incubation period , the slides 45 Oleic acid 10 were washed in PBS ( three times , 5 min each ) , fixed with Trichloromonofluoromethane 5 ,000 Dichlorodifluoromethane 10 ,000 4 % paraformaldehyde in PBS for 10 min in the dark , washed 5 ,000 again with PBS ( three times , 5 min each ) , incubated in the Dichlorotetrafluoroethane dark for 5 min with 300 nM DAPI solution , and mounted ( vii) Topical Gel 1 wt. % with anti - quenching mounting medium . Fluorescence of 50 – " Compound X 5 % FITC was detected with excitation at 460 - 500 nm and Carbomer 934 1 .25 % emission at 512 -542 nm . DAPI was detected with excitation Triethanolamine q . s . at 340 - 380 nm and emission at 425 - 00 nm . (pH adjustment to 5 - 7 ) Statistical Analyses . Methyl paraben 0 . 2 % Data are expressed as mean : SD . Wound healing between 55 Purified water q. s . to 100 g the groups in the studies were analyzed for statistical sig (viii ) Topical Gel 2 wt. % nificance using a paired Student t- test , where p < 0 .05 was considered statistically significant. Compound X ' 5 % Methylcellulose 2 % Methyl paraben 0 . 2 % Example 4 . Pharmaceutical Dosage Forms 60 Propyl paraben 0 .02 % Purified water q . s . to 100 g The following formulations illustrate representative phar maceutical dosage forms that may be used for the therapeu ( ix ) Topical Ointment wt. % tic or prophylactic administration of a compound of a ' Compound X 5 % formula described herein , a compound specifically disclosed 65 Propylene glycol 1 % herein , or a pharmaceutically acceptable salt or solvate Anhydrous ointment base 40 % thereof (hereinafter referred to as “Compound X ' ): US 9 , 951 , 035 B2 67 -continued m Polysorbate 80 2 % Methyl paraben 0 . 2 % HN Purified water q .s . to 100 g ( x ) Topical Cream 1 wt. % "Compound X ' 5 % White bees wax 10 % Liquid paraffin 30 % or a salt thereof. Benzyl alcohol 5 % 10 2 . The compound of claim 1 wherein the stereochemistry Purified water q . s . to 100 g of the thiirane chiral center is in the R configuration . ( xi) Topical Cream 2 wt. % 3. The compound of claim 2 wherein the compound is : 'Compound X 5 % Stearic acid 10 % 15 Glyceryl monostearate 3 % Polyoxyethylene stearyl ether 3 % Sorbitol 5 % Isopropyl palmitate 2 % Methyl Paraben 0 . 2 % Purified water q. s . to 100 g 20 H2N These formulations may be prepared by conventional procedures well known in the pharmaceutical art . It will be (ND -336 , 5 ) appreciated that the above pharmaceutical compositions 25 Lori may be varied according to well- known pharmaceutical techniques to accommodate differing amounts and types of active ingredient 'Compound X ' . Aerosol formulation (vi ) HN may be used in conjunction with a standard , metered dose aerosol dispenser. Additionally , the specific ingredients and 30 proportions are for illustrative purposes . Ingredients may be exchanged for suitable equivalents and proportions may be noun varied , according to the desired properties of the dosage (ND - 378 , 6 ) form of interest. While specific embodiments have been described above 35 with reference to the disclosed embodiments and examples , IZ such embodiments are only illustrative and do not limit the scope of the invention . Changes and modifications can be made in accordance with ordinary skill in the art without morgana departing from the invention in its broader aspects as defined 40 in the following claims. All publications, patents , and patent documents are incor porated by reference herein , as though individually incor NH porated by reference . No limitations inconsistent with this disclosure are to be understood therefrom . The invention has 45 been described with reference to various specific and pre HON ferred embodiments and techniques . However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the con invention . 50 or a pharmaceutically acceptable salt thereof. What is claimed is: 4 . The compound of claim 2 wherein the compound is : 1 . A compound of Formula I : 55 ( R -ND - 336 ) omnimo60 HN wherein 65 or a pharmaceutically acceptable salt thereof. Rl is CH2 NHRCwherein Ra is H or ( C , -C6 ) alkanoyl; 5 . The compound of claim 1 wherein the stereochemistry – NH C ( NH) 2NH2 ; or of the thiirane chiral center is in the S configuration . US 9 , 951, 035 B2 69 70 6 . The compound of claim 5 wherein the compound is : 10 . A composition comprising the compound of claim 4 in combination with a pharmaceutically acceptable diluent , excipient, or carrier. ( 4) 11. A composition comprising the compound of claim 7 in 5 combination with a pharmaceutically acceptable diluent, excipient, or carrier. 12. A method to inhibit MMP -2 , MMP -9 , MMP- 14 , or a combination thereof, comprising contacting a composition HN that includes MMP- 2 , MMP- 9 , or MMP - 14 , with a com S ; 10 pound of claim 1 , thereby inhibiting the enzymatic activity of the matrix metalloproteinase . (ND - 336 , 5 ) 13 . The method of claim 12 wherein the contacting is in vitro . 14 . The method of claim 12 wherein the contacting is in vivo . H2N 15 15 . The method of claim 12 wherein the inhibition is selective for MMP - 2 , MMP - 9 , or both MMP - 2 and MMP - 9 , S ; in the presence of MMP - 8 . mogy(ND -378 , 6 ) 16 . The method of claim 12 wherein the compound is a nanomolar slow - binding inhibitor of MMP - 2 , MMP - 9 , and 20 MMP - 14 , and the compound poorly inhibits MMP - 8 in a non - competitive manner. 17 . A method of accelerating the healing process of a wound comprising administering to a mammal afflicted with a wound an effective amount of a compound of claim 1 , or yhogy 25 a pharmaceutically acceptable salt thereof, wherein the compound is a selective MMP inhibitor and the healing process of the wound is accelerated . NH 18 . The method of claim 17 wherein the compound is

30 H2N IoanZ ( R -ND -336 , 5 ) or a pharmaceutically acceptable salt thereof. 35 H2N y av7 . iThe romantic compound love of claim you 5 wherein themore compoundompound on isdess. :w a ( S - ND - 336 ) or a pharmaceutically acceptable salt thereof. 40 19 . The method of claim 17 wherein the compound is

HN ( S -ND - 336 , 5 ) S ; H 45 or a pharmaceutically acceptable salt thereof. HN 8 . A composition comprising the compound of claim 1 in combination with a pharmaceutically acceptable diluent, 50 H excipient, oroon carrier . on 9. The composition of claim 8 wherein the composition is or a pharmaceutically acceptable salt thereof. a topical formulation . * * * * *